sign in sign up
<<
Home , Melatonin receptor 1C, Neuromedin U receptor 1, Neurotensin receptor 2, OPN1SW, Somatostatin receptor 5, Thyrotropin receptor

cDNA Resource Center

cDNA Resource Center

Catalog

cDNA Resource Center Missouri University of Science and Technology 400 W 11th Rolla, MO 65409

TEL: (573) 341-7610 FAX: (573) 341-7609 EMAIL: [email protected]

www.cdna.org

September, 2008

1 cDNA Resource Center

Visit our web site for product updates

2 cDNA Resource Center

The cDNA Resource Center

The cDNA Resource Center is a service provided by the faculty of the Department of Biological Sciences of Missouri University of Science and Technology.

The purpose of the cDNA Resource Center is to further scientific investigation by providing cDNA clones of involved in signal transduction processes. This is achieved by providing high quality clones for important signaling proteins in a timely manner.

By high quality, we mean that the clones are • Sequence verified • Propagated in a versatile vector useful in bacterial and mammalian systems • Free of extraneous 3' and 5' untranslated regions • Expression verified (in most cases) by coupled in vitro transcription/translation assays • Available in wild-type, epitope-tagged and common mutant forms (e.g., constitutively- active or dominant negative)

By timely, we mean that the clones are • Usually shipped within a day from when you place your order.

Clones can be ordered from our web pages, by FAX or by phone. Within the United States, clones are shipped by overnight courier (FedEx); international orders are shipped International Priority (FedEx).

The clones are supplied for research purposes only. Details on use of the material are included on the Material Transfer Agreement (page 3).

Clones are distributed by agreement in Invitrogen's pcDNA3.1+ vector. The clones are provided as dried DNA (approximately 1.0 µg) on a circle of Whatman #1 filter paper. Instructions for reconstituting the DNA are provided in the FAQ section.

Questions can be directed toward any Resource Center staff member at (573) 341-7610, Monday through Friday between 8:30 AM-4:30 PM CT. You may also contact us by FAX at (573) 341-7609, or email at [email protected].

For more information, refer to the FAQ section beginning on page 4.

3 cDNA Resource Center

MATERIAL TRANSFER AGREEMENT FOR cDNA CLONES

Acceptance of any of the clones or associated products (hereinafter “Material”) provided by the cDNA Resource Center is contingent upon your good faith agreement to the following terms.

1. You will use the Material only for research purposes to advance the public interest. This Material is provided for use only in laboratory animals or in vitro experiments; the provided Material will not be used in human beings.

2. You agree that the clones are being provided without warranty, express or implied, including any implied warranty of merchantability or fitness for a particular purpose or freedom from infringement of any patent or other proprietary right of a third party.

3. You agree that these terms apply to purchases of Material: • A fee of $120 is made for each clone purchased. • Credit cards (MasterCard or Visa) or purchase orders are to be used exclusively for purchases with payment terms of net 30 for purchase orders. • FOB shipping point. A charge to purchaser of $15.50 is added to the cost of each shipment for overnight delivery in the US and $40.00 for international priority delivery.

4. You understand that third parties may claim proprietary rights to the commercial use of the Material provided. This agreement does not constitute a waiver of any patent rights.

5. You agree to hold harmless and indemnify the Curators of the Missouri University of Science and Technology, the provider(s) of the Material and persons acting on their behalf, for any claim asserted by a third party related to your possession, use, storage, or disposal of the Material.

6. You will not redistribute the Material except by special agreement with the cDNA Resource Center. In addition, you will not use the provided Material in a manner that duplicates or undermines the efforts of the cDNA Resource Center.

7. In scientific communications describing work utilizing the provided Material, you will acknowledge the source of the Material as being the cDNA Resource Center, referencing our Web Site (www.cdna.org ).

4 cDNA Resource Center

Frequently Asked Questions

1. What is the cDNA Resource Center? the of interest. The sequence is then amplified through PCR as above. Regardless of the source The cDNA Resource Center is a service provided the ORF is amplified using a high fidelity by the faculty of the Department of Biological polymerase (e.g., Pfu). Sciences of the Missouri University of Science and Technology. In both cases, the PCR product is then subcloned into a plasmid vector (see below) and then sequence The purpose of the cDNA Resource Center is to verified against published sequences. further scientific investigation by providing cDNA Discrepancies, if any, from the published clones of human proteins involved in signal sequence(s) are noted in the clone documentation. transduction processes. The goal of the Center is to All wildtype sequences are then submitted to accelerate scientific discovery in human biology GenBank to catalog the coding sequences (CDS) and pathology. This is achieved by providing high resolved through clone production. quality clones for important signaling proteins in a timely manner. We have also obtained clones from investigators who wish to deposit their clones in the Resource By high quality, we mean that the clones are Center (e.g., WOX1, AGS1). The ORF of deposited • Sequence verified clones are subcloned into the standard vector and • Propagated in a versatile vector the sequence of the insert is verified. We thus (pcDNA3.1+) maintain standard "contexts" for each clone in the • Free of extraneous 3' and 5' untranslated collection. regions • Available in wild-type, epitope-tagged 3. Which vector are the clones in? and common mutant forms (e.g., constitutively- active or dominant negative) Clones are distributed by agreement in Invitrogen's • Expression-verified in most cases by pcDNA3.1+ vector. coupled in vitro transcription/translation assays By timely, we mean that the clones are pcDNA3.1+ • Usually shipped within a day of order by This vector can be used for mammalian courier delivery (Federal Express). expression (CMV promoter, bovine polyadenylation signal), production of We expect this service to allow scientists to focus sense strand RNA by in vitro transcription (T7), on basic biological questions without spending production of single stranded DNA (f1 origin), and excessive amounts of time in procuring, preparing production of stably transfected cell lines or verifying their own (or their colleagues) cDNA (neomycin resistance). The vector contains the clones. We expect these clones to be particularly ampicillin resistance gene and can be propagated in useful to scientists probing and reconstructing common E. coli strains. (www.invitrogen.com) signal transduction pathways in model systems. 4. How do I order the clones?

2. How are the clones obtained? Clones can be ordered via our web page (www.cdna.org), by FAX (573 341-7609) or by At present, the clones are obtained in one of two telephone (573-341-7610). Purchase order (P.O.) ways: Most frequently, the polymerase chain numbers from research organizations based in the reaction (PCR) is used to amplify the ORF from United States are accepted. Credit card (MasterCard commercially prepared cDNA. Alternatively, a or Visa) orders are accepted from all research computer search of GenBank human reference organizations (US and international). sequences and other web-tools are used to identify potential genomic (intron-free) sequences as s source for complete open reading frame (ORF) of

5 cDNA Resource Center

5. How are the clones shipped? OPENING to ensure pellet is at the bottom of the tube. Add 50ul of ddH20 or TE buffer (as above) All orders are generally shipped within a day of and vortex or flick the tube to ensure that the pellet being placed and are sent via courier delivery dissolves. (Federal Express). For Both: Use 1-2 µl of supernatant for use in The clones are provided in one of two ways; as transfecting E. coli by electroporation or chemical dried DNA (approximately 1.0 µg) on a circle of means. Please do not try to use the DNA directly Whatman #1 filter paper, or, with increasing for any application other than to transform bacteria frequency, as dried DNA (approximately 2.0 µg) and prepare a large stock. within a 0.5 ml skirted tube. Instructions for reconstituting the DNA are provided below. 8. What are the restrictions on the use of the clones? 6. Documentation The clones are supplied solely for research For each clone, up to four forms of documentation purposes. The clones are distributed with the can be downloaded from our web site: 1) an understanding that you will not redistribute them to information sheet that contains details on the insert, others. Details on use of the material are included including how the clone was obtained, any on the Material Transfer Agreement that is sequence discrepancies, and experimental available on this web site. mutagenesis details, if applicable; 2) a sequence sheet that contains the nucleotide sequence of the 9. How should I acknowledge the source of the insert and the predicted translation product; 3) a text clones in manuscripts? file in FASTA format containing the insert nucleotide sequence can be downloaded for use Scientific communications describing work with your molecular biology software (this usually utilizing clones or associated material provided by contains small regions of the flanking plasmid the cDNA Resource Center Material should sequence as well); 4) for the relevant clones a PDF acknowledge this source, referencing our Web Site. file documenting expression in a coupled For example: "The cDNA clone for human Go- transcription/translation assay system. PDF files can alpha was obtained from the cDNA Resource be downloaded and viewed with Adobe Acrobat Center (www.cdna.org)." Reader (www.adobe.com). 10. Who do I contact regarding shipping and/or 7. How do I "reconstitute" the cDNA after it technical issues? arrives? Questions can be directed toward any Resource Store clones at 4°C until you are ready to use Center staff member at (573) 341-7610 on Monday them. Clones are guaranteed for up to 6 months through Friday between 8:30 AM-4:30 PM, CT. We from the date purchased. Clones are supplied as may also be contacted by FAX at (573) 341-7609, DNA (approximately 1.0 µg) spotted onto either a or email at [email protected]. Whatman #1 filter disks, or dried within a 0.5ml screw-cap skirted tube. 11. Who can I contact with suggestions for improving the Resource Center? Whatman Disk: Add 100 µl of TE buffer (10 mM TRIS base, 1 mM EDTA, pH 8.0) to the micro- Suggestions and/or comments regarding the cDNA centrifuge tube w/ filter paper, vortex briefly, Resource Center including suggestions for incubate at room temperature for 5 minutes, and improvement and future directions can be directed repeat the vortex. Centrifuge the tube for a few toward any Center staff member: (TEL) 573 341- seconds and then remove the filter paper from the 7610; (FAX) 573 341-7609; email ([email protected]). tube.

0.5ml Tube: Centrifuge the tube BEFORE

6 cDNA Resource Center Clone Collection GPCRs 7 Y 17 FLAG tagged 33 Acetylcholine (Muscarinic) 7 18 PDNR-1r 33 7 Nucleotides/P2Y 18 © subunits 35 Adrenergic 8 Opioid 19 Wild-Type 35 8 19 HA tagged 35 Angiotensin 8 Orphan 19 PDNR-1r9 36 Arginine Vasopressin 9 Oxytocin 20 9 20 G Proteins Regulators 38 9 Platelet-Activating Factor 20 AGS (activators of G 9 Prokineticin 20 signaling) 38 Cannabinoid 9 Releasing RCC1 38 Chemokine 10 Hormone 20 RGS (regulators of G protein 12 Prostaglandins and signaling) 38 Coagulation Thromboxane 20 Rho GDI (guanine nucletide Factor/Thrombin 12 /-Like 21 dissociation inhibitor) 39 Complement component 5a 21 (C5a) 12 Serotonin/5-HT 21 Small G-Proteins 40 Corticotropin Releasing 22 ARF family 40 Hormone 12 Succinate 23 Wild-Type 40 Cysteinyl Leukotriene 12 Tachykinin 23 PDNR-1r 40 Dopamine 13 Thyroid Stimulating Rab family 41 13 Hormone 23 Wild-Type 41 Estroogen 13 Thyrotropin-Releasing PDNR-1r 43 Follicle Stimulating Hormone13 Hormone 23 Ran family 44 Formyl Peptide 13 Trace Amine 23 Wild-Type 44 Free Fatty Acid 14 Transmembrane 7 3xHA tagged 44 Homolog 14 Superfamily 23 Ras family 45 14 Urotensin II 23 Wild-Type 45 14 Vasoactive Intestinal Peptide24 pDNR-1r 45 Glutamate 14 GV Mutants 46 Gonadotropin-Releasing GPCR Regulators 25 3xHA GV 46 Hormone 14 25 SN Mutants 47 Growth Hormone Releasing GRB Protein 25 3xHA SN 47 Hormone 15 RAMP Proteins 25 3xHA-Tagged 47 Growth Hormone 2XMYC GV 48 Secretagogue 15 Heterotrimeric G-Proteins 2XMYC SN 48 Histamine 15 〈 subunits 26 2XMYC-tagged 48 Hypocretin 15 Wild-Type 26 Ras-like 49 KISS 15 PDNR-1r 27 Wild-Type 49 -Rich Repeat PTX-resistant Mutants 28 PDNR-1r 49 Containing 15 QL mutants (constitutively Rho family 50 15 active) 29 Wild-Type 50 /S1P 15 XTP mutants (dominant pDNR-1r 51 MAS –Related 16 negative) 29 3xHA-Tagged 52 Melanin-Concentrating EE-tagged (internal tag) 30 GV Mutants 52 Hormone 16 GT mutants (dominant 3xHA GV Mutants 52 Melanocortin 17 negative) 31 TN Mutants 53 17 QL/EE tagged 3xHA TN Mutants 53 17 constitutively active 31 Other Rho mutants 54 17 ® subunits 33 Neuropeptide FF 17 Wild-Type 33

7 cDNA Resource Center

G Protein Coupled Receptors

Acetylcholine (Muscarinic) Leukotriene B4 Adenosine Lysophosphatidic Acid/S1P Adrenergic MAS-related Adrenomedullin Melanin-Concentrating Hormone Angiotensin Melanocortin Arginine Vasopressin Melatonin Bombesin Motilin Bradykinin Neuromedin U Calcitonin Neuropeptide FF Cannabinoid Chemokine Neurotensin Cholecystokinin Nucleotides/P2Y Coagulation Factor II Opioid Complement component 5a (C5a) Opsin Corticotropin Releasing Hormone Orphan Receptors Cysteinyl Leukotriene Oxytocin Dopamine Parathyroid Hormone Endothelin Platelet-Activating Factor Prokineticin Follicle Stimulating Hormone Prolactin Releasing Hormone Formyl Peptide Prostaglandins and Thromboxanes Free Fatty Acid Relaxin/Insulin-Like Peptide Frizzled Homolog Secretin Galanin Serotonin/5-HT Glucagon Somatostatin Glutamate (Metabotropic) Succinate Gonadotropin-Releasing Hormone Tachykinin Growth Hormone Releasing Hormone Thyroid stimulating hormone Growth Hormone Secretagogue Thyrotropin-Releasing Hormone Histamine Trace Amine Hypocretin Transmembrane 7 Superfamily KISS Receptors Urotensin-II Leucine-Rich Repeat-Containing Vasoactive Intestinal Peptide

Muscarinic Receptors acetylcholine M4 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho I (3'). The open reading MAR0100000 Muscarinic 1 (M1) frame was amplified by the PCR from human whole brain cDNA (a.k.a: muscarinic acetylcholine receptor M1). GenBank (Clontech). The insert was sequenced. Insert size = 1450 bp. Accession # AF498915: Human muscarinic acetylcholine MAR040TN00 Muscarinic acetylcholine receptor 4 (M4) 3xHA- receptor M1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at tagged(N-terminus) BamH I (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA (Clontech). MAR0500000 Muscarinic acetylcholine receptor 5 (M5) The insert was sequenced. Insert size = 1390 bp. GenBank Accession # AF498919: Human muscarinic MAR010TN00 Muscarinic acetylcholine receptor 1 (M1) 3xHA- acetylcholine receptor M5 (wild type) cloned into pcDNA3.1+ tagged(N-terminus) (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA MAR0200000 Muscarinic acetylcholine receptor 2 (M2) (Clontech). The insert was sequenced. Insert size = 1610 bp. GenBank Accession # AF498916: Human muscarinic MAR050TN00 Muscarinic acetylcholine receptor 5 (M5) 3xHA- acetylcholine receptor M2 (wild type) cloned into pcDNA3.1+ tagged(N-terminus) (Invitrogen) at EcoR I (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 1410 bp. Adenosine Receptors MAR020TN00 Muscarinic acetylcholine receptor 2 (M2) 3xHA-tagged(N-terminus) ADRA100000 A1 (ADORA1) (Alternate Symbols: RDC7). GenBank Accession # AY136746: MAR0300000 Muscarinic acetylcholine receptor 3 (M3) Human adenosine receptor ADORA1 (wild type) cloned into GenBank Accession # AF498917: Human muscarinic pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open acetylcholine receptor M3 (wild type) cloned into pcDNA3.1+ reading frame was amplified by the PCR from human whole (Invitrogen) at BamH I (5') and Xba I (3'). The open reading brain cDNA (Clontech). The insert was sequenced. Insert size = frame was amplified by the PCR from human whole brain cDNA 980 bp. (Clontech). The insert was sequenced. Insert size = 1790 bp. ADRA10TN00 Adenosine receptor A1 (ADORA1) 3xHA-tagged MAR030TN00 Muscarinic acetylcholine receptor 3 (M3) (N-terminus) 3xHA-tagged(N-terminus) ADRA2A0000 Adenosine Receptor A2A (ADORA2A) MAR0400000 Muscarinic acetylcholine receptor 4 (M4) (Alternate Symbols: RDC8, hA2aR, ADORA2); (a.k.a: GenBank Accession # AF498918: Human muscarinic adenosine A2 receptor; adenosine receptor subtype A2a).

8 cDNA Resource Center

GenBank Accession # AY136747: Human adenosine receptor ADORA2A (wild type) cloned into pcDNA3.1+ (Invitrogen) at AR0A2C0000 alpha 2C (ADRA2C) BamHI (5') and XhoI (3'). The open reading frame was amplified (Alternate Symbols: ADRA2L2, ADRARL2, ADRA2RL2); (a.k.a: by the PCR from IMAGE clone (4385637). The insert was alpha2-AR-C4). Genbank ACC# AY455666: Human adrenergic sequenced. Insert size = 1240 bp. receptor ADRA2C (wild type) cloned into pcDNA3.1+ (Invitrogen) ADRA2ATN00 Adenosine receptor A2A (ADORA2A) 3xHA- at BamHI (5') and XhoI (3'). The open reading frame was tagged (N-terminus) amplified by the PCR from human genomic DNA. The insert was sequenced. Insert size = 1377 bp. ADRA2B0000 Adenosine Receptor A2B (ADORA2B) (Alternate Symbols: ADORA2). GenBank Accession # AR0B200000 Adrenergic Receptor beta 2 (ADRB2) AY136748: Human adenosine receptor ADORA2B (wild type) (Alternate Symbols: BAR, B2AR, ADRBR, ADRB2R); (a.k.a: cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XbaI (3'). beta-2 adrenoceptor; catecholamine receptor; beta-2 adrenergic The open reading frame was amplified by the PCR from human receptor). Genebank ACC# NM_000024.3: Human adrenergic whole brain cDNA (Clontech). The insert was sequenced. Insert receptor ADRB2 (wild type) cloned into pcDNA3.1+ (Invitrogen) size = 1000 bp. Stop codon with adjacent XbaI site form DAM at EcoRI (5') and XhoI (3'). The open reading frame was methylation sensitive site. amplified by the PCR from human heart cDNA (clontech). The ADRA2BTN00 Adenosine receptor A2B (ADORA2B) 3xHA- insert was sequenced. Insert size = 1250 bp. tagged (N-terminus) AR0B20TN00 Adrenergic receptor beta 2 (ADRB2) 3xHA-tagged (N-terminus) ADRA300000 Adenosine Receptor A3 (ADORA3) (Alternate Symbols: A3AR). GenBank Accession # AY136749: AR0B300000 Adrenergic receptor beta 3 (ADRB3) Human adenosine receptor ADORA3 (wild type) cloned into Genbank ACC# AY487247: Human adrenergic receptor beta 3 pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open (ADRB3) (wild type) cloned into pcDNA3.1+ (Invitrogen) at reading frame was amplified by the PCR from human whole BamHI (5') and XhoI (3'). The open reading frame was amplified brain cDNA (Clontech). The insert was sequenced. Insert size = by the PCR from human placental cDNA (Clontech). The insert 960 bp was sequenced. Insert size = 1227 bp. ADRA30TN00 Adenosine receptor A3 (ADORA3) 3xHA-tagged (N-terminus) Adrenomedullin receptor Adrenergic Receptors ADMR000000 Adrenomedullin receptor (ADMR) AR0A1A0000 Adrenergic Receptor alpha 1A (ADRA1A) GenBank ACC# AY268434: Adrenomedullin receptor (ADMR) (Alternate Symbols: ADRA1C, ADRA1L1); (a.k.a: adrenergic, (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and alpha-1C-, receptor; adrenergic, alpha -1A-, receptor). Genbank XhoI (3'). The open reading frame was amplified by the PCR ACC# AY389505: Human adrenergic receptor ADRA1A (wild from Human Genomic DNA . The insert was sequenced. Insert type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI size= 1215bp N.B. This gene encodes a seven transmembrane (3'). The open reading frame was amplified by the PCR from receptor that shares sequence similarity with the rat human heart cDNA (Clontech). The insert was sequenced. Insert adrenomedullin receptor. This similarity suggests that potential size = 1401 bp. ligands of the encoded protein will be members of the calcitonin AR0A1ATN00 Adrenergic receptor alpha 1A (ADRA1A) 3xHA- gene-related peptide// adrenomedullin peptide family. As tagged (N-terminus) noted previously (Kennedy SP, Sun D. Oleynek JJ, Hoth CF, Kong J, Hill RJ. 1998. "Expression of the rat adrenomedullin AR0A1B0000 Adrenergic receptor alpha 1B (ADRA1B) receptor or a putative human adrenomedullin receptor does not (Alternate Symbols: ADRA1); (a.k.a: adrenergic, alpha -1B-, correlate with adrenomedullin binding or functional response". receptor). Identical to Genbank ACC# NM_000679 with the Biochim. Biophys. Res. Commun. 244:832-837) it is likely that following exception G549->A (Silent mutation): Human this putative GPCR remains an orphan, as the ability to bind and adrenergic receptor alpha 1B (ADRA1B) (wild type) cloned into be activated by adrenomedullin has been refuted. A complex of pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open like-receptor (CRLR) and RAMP2 or RAMP3 reading frame was amplified by the PCR from human whole is believed to give a functional receptor. brain cDNA (Clontech). The insert was sequenced. Insert size = ADMR00TN00 Adrenomedullin receptor (ADMR) 3xHA-tagged 1563 bp. (N-terminus)

AR0A2A0000 Adrenergic Receptor alpha 2A (ADRA2A) (Alternate Symbols: ADRA2, ADRAR, ZNF32, ADRA2R); (a.k.a: Angiotensin Receptor alpha-2AAR subtype C10). Identical to the Genbank ACC# NM_000681 with the following exeption (G855A) silent mutation: AGTL100000 Angiotensin receptor-like 1 (AGTRL1) Human adrenergic receptor ADRA2A (wild type) cloned into (Alternate Symbols: APJ, MGC45246); (a.k.a: angiotensin pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open receptor-like 1). Identical to GenBank ACC# NM_005161 with reading frame was amplified by the PCR from human genomic the following exeption (C135A) (Silent mutation): angiotensin DNA. The insert was sequenced. Insert size = 1353 bp. receptor-like 1 (AGTRL1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame AR0A2B0000 Adrenergic Receptor alpha 2B (ADRA2B) was amplified by the PCR from Human Genomic DNA . The (Alternate Symbols: ADRA2L1, ADRARL1, ADRA2RL1); (a.k.a: insert was sequenced. Insert size= 1143bp alpha-2-adrenergic receptor-like 1). Genbank ACC# AY548167: AGTL10TN00 Angiotensin receptor-like 1 (AGTRL1) 3xHA- Human adrenergic receptor ADRA2B (wild type) cloned into tagged (N-terminus) pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human genomic AGTR100000 angiotensin II receptor (Variant 1) AGTR1 DNA. Insert size = 1353 bp. (Alternate Symbols: AT1, AG2S, AT1B, AT2R1, HAT1R, AR0A2BTN00 Adrenergic receptor alpha 2B (ADRA2B) 3xHA- AGTR1A, AGTR1B, AT2R1A, AT2R1B); (a.k.a: angiotensin tagged (N-terminus)

9 cDNA Resource Center receptor 1; angiotensin receptor 1B; type-1B angiotensin II Bradykinin Receptors receptor). Identical to GenBank ACC# NM_000685: angiotensin II receptor (Variant 1) AGTR1 (wild type) cloned into pcDNA3.1+ BDKB100000 B1 (BDKRB1) (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame GenBank ACC# AY275464: (BDKRB1) was amplified by the PCR from Human Genomic DNA . The (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and insert was sequenced. Insert size= 1080 bp XhoI (3'). The open reading frame was amplified by the PCR AGTR10TN00 Angiotensin II receptor (Variant 1) AGTR1 3xHA- from human genomic DNA . The insert was sequenced. Insert tagged (N-terminus) size= 1062 bp BDKB10TN00 Bradykinin receptor B1 (BDKRB1) 3xHA-tagged AGTR200000 Angiotensin II receptor, type 2 (AGTR2) (N-terminus) (Alternate Symbols: ATR2L1); (a.k.a: angiotensin receptor-like 2; angiotensin II receptor-like). Identical to GenBank ACC# BDKB200000 (BDKRB2) AY322550: Angiotensin II receptor, type 2 (AGTR2) (wild type) GenBank ACC# AY275465: Bradykinin receptor B2 (BDKRB2) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and The open reading frame was amplified by the PCR from Human XhoI (3'). The open reading frame was amplified by the PCR Genomic DNA . The insert was sequenced. Insert size= 1092bp from human lung cDNA . The insert was sequenced. Insert size= AGTR20TN00 Angiotensin II receptor, type 2 (AGTR2) 3xHA- 1176 bp tagged (N-terminus) BDKB20TN00 Bradykinin receptor B2 (BDKRB2) 3xHA-tagged (N-terminus)

Arginine Vasopressin Receptors Calcitonin Receptors AVR01A0000 Arginine 1A (AVPR1A) (Alternate Symbols: AVPR1); (a.k.a: V1a vasopressin receptor; CALCR00000 Calcitonin receptor (CALCR) antidiuretic 1A; SCCL vasopressin subtype 1a (Alternate Symbols: CRT, CTR, CTR1). GenBank ACC# receptor; V1-vascular vasopressin receptor AVPR1A; AY430048: Calcitonin receptor (CALCR) (wild type) cloned into vascular/hepatic-type arginine vasopressin receptor). GenBank pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI (3'). The open ACC# AY322550: Arginine (AVPR1A) reading frame was amplified by the PCR from human kidney (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and cDNA (Clontech) . The insert was sequenced. Insert size= 1425 XhoI (3'). The open reading frame was amplified by the PCR bp. from human liver cDNA (Clontech). Insert size= 1257bp. AVR01ATN00 Arginine vasopressin receptor 1A (AVPR1A) CALCRL0000 Calcitonin receptor-like (CALCRL) 3xHA-tagged (N-terminus) (Alternate Symbols: CRLR, CGRPR). GenBank ACC# AY389506: Calcitonin receptor-like (CALCRL) (wild type) cloned AVR0200000 Arginine (AVPR2) into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open (nephrogenic diabetes insipidus) reading frame was amplified by the PCR from human heart (Alternate Symbols: DI1, DIR, NDI, V2R, ADHR). GenBank cDNA (Clontech) . The insert was sequenced. Insert size= 1386 ACC# AY242131: Arginine vasopressin receptor 2 (AVPR2) (wild bp type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI CALCRLTN00 Calcitonin receptor-like (CALCRL) 3xHA-tagged (3'). The open reading frame was amplified by the PCR from (N-terminus) kidney cDNA. Insert size= 1116 bp. AVR020TN00 Arginine vasopressin receptor 2 (AVPR2) 3xHA- tagged (N-terminus) Cannabinoid Receptors

CNR01L0000 1 (long isoform) (CNR1L) Bombesin Receptors (Alternate Symbols: CB1, CNR, CB-R, CB1A, CANN6, CB1K5); (a.k.a: central cannabinoid receptor). GenBank ACC# BRS0300000 Bombesin-like receptor 3 (BRS3) AY225225: cannabinoid receptor 1 (long isoform) (CNR1L) (wild GenBank ACC# AY585193: Bombesin-like receptor 3 (BRS3) type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (3'). The open reading frame was amplified by the PCR from XhoI (3'). The open reading frame was amplified by the PCR Human Genomic DNA . The insert was sequenced. Insert size= from Human testis cDNA . The insert was sequenced. Insert 1419 bp size= 1200bp. CNR01LTN00 Cannabinoid receptor 1 (long isoform) (CNR1L) BRS030TN00 Bombesin-like receptor 3 (BRS3) 3xHA-tagged 3xHA-tagged (N-terminus) (N-terminus) CNR0200000 Cannabinoid receptor 2 (CNR2) GRPR000000 Releasing Peptide Receptor (GRPR) (Alternate Symbols: CB2, CX5). GenBank ACC# AY242132: Identical to GenBank ACC# NM_005314 with the exeption of two Cannabinoid receptor 2 (CNR2) (wild type) cloned into silent mutations (C453T) and (C663T) : Gastrin releasing peptide pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open receptor (GRPR) (wild type) cloned into pcDNA3.1+ (Invitrogen) reading frame was amplified by the PCR from human genomic at EcoRI (5') and XhoI (3'). The open reading frame was DNA. Insert size= 1083bp. amplified by the PCR from Human brain cDNA (Clontech) . The CNR020TN00 Cannabinoid receptor 2 (CNR2) 3xHA-tagged (N- insert was sequenced. Insert size= 1155bp terminus) GRPR00TN00 Gastrin-releasing peptide receptor (GRPR) 3xHA- tagged

10 cDNA Resource Center

Chemokine Receptors CCR050TN00 Chemokine (C-C motif) receptor 5 (CCR5) 3xHA- tagged (N-terminus) CCBP200000 Chemokine binding protein 2 (CCBP2) (Alternate Symbols: D6, CCR9, CCR10, CMKBR9); (a.k.a: hD6; CCR0600000 Chemokine (C-C motif) receptor 6 (CCR6) chemokine (C-C) receptor 9; chemokine (C-C motif) receptor 9; (Alternate Symbols: BN-1, CKR6, DCR2, CKRL3, DRY-6, CC-chemokine-binding receptor JAB61). GenBank ACC# GPR29, CKR-L3, CMKBR6, GPRCY4, STRL22, GPR-CY4); AY262687: Chemokine binding protein 2 (CCBP2) (wild type) (a.k.a: -like 3; chemokine (C-C) receptor 6; G cloned into pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI (3'). protein-coupled receptor 29; seven-transmembrane receptor, The open reading frame was amplified by the PCR from genomic lymphocyte, 22). GenBank ACC# AY242126: Chemokine (C-C DNA. Insert size= 1155 bps. motif) receptor 6 (CCR6) (wild type) cloned into pcDNA3.1+ CCBP20TN00 Chemokine binding protein 2 (CCBP2) 3xHA- (Invitrogen) at BamHI(5') and XhoI (3'). The open reading frame tagged (N-terminus) was amplified by the PCR from leukocyte cDNA. Insert size= 1125 bp. CCR0100000 Chemokine (C-C motif) receptor 1 (CCR1) CCR060TN00 Chemokine (C-C motif) receptor 6(CCR6) 3xHA- (Alternate Symbols: CKR-1, HM145, CMKBR1, MIP1aR); (a.k.a: tagged (N-terminus) RANTES receptor). Chemokine (C-C motif) receptor 1 (CCR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI(5') and CCR0700000 Chemokine (C-C motif) receptor 7 (CCR7) XhoI (3'). The open reading frame was amplified by the PCR GenBank ACC# AY587876: Chemokine (C-C motif) receptor 7 from a clone obtained from the Laboratory of Genetics/National (CCR7) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI Institutes of Mental Health at the NIH. The insert was sequenced (5') and XhoI (3'). The open reading frame was amplified by the and found to be identical to GenBank ACC# NM_001295. Insert PCR from Human Leukocyte cDNA . The insert was sequenced. size= 1068 bp. Insert size= 1137bp. CCR010TN00 Chemokine (C-C motif) receptor 1 (CCR1) 3xHA- CCR070TN00 Chemokine (C-C motif) receptor 7(CCR6) 3xHA- tagged (N-terminus) tagged (N-terminus)

CCR0200000 Chemokine (C-C motif) receptor 2 (CCR2) isoform CCR0800000 Chemokine (C-C motif) receptor 8 (CCR8) A (Alternate symbols: CY6, TER1, CKRL1, CKR-L1, CMKBR8, (Alternate Symbols: CKR2, CCR2A, CCR2B, CKR2A, CKR2B, CMKBRL2, GPR-CY6); (a.k.a: chemokine (C-C) receptor 8; CC- CMKBR2, MCP-1-R, CC-CKR-2); (a.k.a: chemokine (C-C) chemokine receptor chemr1; chemokine (C-C) receptor-like 2). receptor 2; monocyte chemotactic protein 1 receptor; monocyte Chemokine (C-C motif) receptor 8 (CCR8) (wild type) cloned into chemoattractant protein 1 receptor). Chemokine (C-C motif) pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open receptor 2 (CCR2) isoform A (wild type) cloned into pcDNA3.1+ reading frame was amplified by the PCR from a clone obtained (Invitrogen) at BamHI(5') and XhoI (3'). The open reading frame from the Laboratory of Genetics/National Institutes of Mental was amplified by the PCR from a clone obtained from the Health at the NIH. The insert was sequenced and found to be Laboratory of Genetics/National Institutes of Mental Health at the identical to GenBank ACC# NM_005201. Insert size= 1068bp. NIH. The insert was sequenced and found to be identical to CCR080TN00 Chemokine (C-C motif) receptor 8 (CCR8) 3xHA- GenBank ACC# AF545480. Insert size= 1083 bp. tagged (N-terminus) CCR020TN00 Chemokine (C-C motif) receptor 2 (CCR2) 3xHA- tagged (N-terminus) CCR0900000 Chemokine (C-C motif) receptor 9 (CCR9) (Alternate Symbols: GPR-9-6). GenBank ACC# AY242127: CCR0300000 Chemokine (C-C motif) receptor 3 (CCR3) Chemokine (C-C motif) receptor 9 (CCR9) (wild type) cloned into (Alternate Symbols: CKR3, CMKBR3, CC-CKR-3); (a.k.a: b- pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open chemokine receptor; eosinophil eotaxin receptor; eosinophil CC reading frame was amplified by the PCR from genomic DNA. chemokine receptor 3). Identical to GenBank ACC# NM_001837: Insert size= 1074 bp. Chemokine (C-C motif) receptor 3 (CCR3) (wild type) cloned into CCR090TN00 Chemokine (C-C motif) receptor 9 (CCR9) 3xHA- pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open tagged (N-terminus) reading frame was amplified by the PCR from Human Genomic DNA . The insert was sequenced. Insert size= 1201bp CCR1000000 Chemokine (C-C motif) Receptor 10 (CCR10) CCR030TN00 Chemokine (C-C motif) receptor 3 (CCR3) 3xHA- (Alternate Symbols: GPR2); (a.k.a: CC chemokine receptor 10). tagged (N-terminus) GenBank ACC# AY429103: Chemokine (C-C motif) receptor 10 (CCR10) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI CCR0400000 Chemokine (C-C motif) receptor 4 (CCR4) (5') and XhoI (3'). The open reading frame was amplified by the (Alternate Symbols: CKR4, k5-5, CMKBR4, ChemR13, CC-CKR- PCR from Human leukocytes cDNA (Clontech) . The insert was 4, HGCN:14099); (a.k.a: chemokine (C-C) receptor 4). Identical sequenced. Insert size= 1089bp to GenBank ACC# AY322539: Chemokine (C-C motif) receptor 4 CCR100TN00 Chemokine (C-C motif) receptor 10 (CCR10) (CCR4) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI 3xHA-tagged (N-terminus) (5') and XhoI (3'). The open reading frame was amplified by the PCR from Human Genomic DNA . The insert was sequenced. CCRL100000 Chemokine (C-C motif) receptor-like 1 (CCRL1) Insert size= 1083bp (Alternate Symbol: PPR1, CCBP2, CCR10, CCR11, VSHK1, CCR040TN00 Chemokine (C-C motif) receptor 4 (CCR4) 3xHA- CKR-11, CCX-CKR, CC-CKR-11); (a.k.a: C-C chemokine tagged (N-terminus) receptor type 11; chemocentryx chemokine receptor; chemokine, cc motif, receptor-like protein 1; orphan seven-transmembrane CCR0500000 Chemokine (C-C motif) receptor 5 (CCR5) receptor, chemokine related). GenBank ACC# AY221094: (Alternate Symbols: CKR5, CKR-5, CCCKR5, CMKBR5, CC- chemokine (C-C motif) recetor-like 1 (CCRL1) (wild type) cloned CKR-5); (a.k.a: chemr13; chemokine (C-C) receptor 5). Identical into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The to GenBank ACC# BC038398: Chemokine (C-C motif) receptor 5 open reading frame was amplified by the PCR from Human (CCR5) (wild type) cloned into pcDNA3.1+ (Invitrogen) at Genomic DNA . The insert was sequenced. Insert size= 1053 bp BamHI(5') and XhoI (3'). The open reading frame was amplified CCRL10TN00 Chemokine (C-C motif) receptor-like1 (CCRL1) by the PCR from Human Genomic DNA . The insert was 3xHA-tagged (N-terminus) sequenced. Insert size= 1059bp

11 cDNA Resource Center

CCRL200000 Chemokine (C-C motif) receptor-like 2 (CCRL2) (Alternate Symbols: HCR, CKRX, CRAM-A, CRAM-B). GenBank CXCR300000 Chemokine (C-X-C motif) receptor 3 (CXCR3) ACC# AY337001: chemokine (C-C motif) recetor-like 2 (CCRL2) (Alternate Symbols: GPR9, IP10, MigR, CD183, Mig-R, CKR-L2, (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and CMKAR3, IP10-R); (a.k.a: Mig receptor; IP10 receptor; G XhoI (3'). The open reading frame was amplified by the PCR protein-coupled receptor 9; chemokine (C-X-C) receptor 3). from Human Lung cDNA (Clontech). The insert was sequenced. GenBank ACC# AY242128: Chemokine (C-X-C motif) receptor 3 Insert size= 1035 bp (CXCR3) (wild type) cloned into pcDNA3.1+ (Invitrogen) at CCRL20TN00 Chemokine (C-C motif) receptor-like2 (CCRL2) EcoRI(5') and XhoI (3'). The open reading frame was amplified 3xHA-tagged (N-terminus) by the PCR from leukocyte cDNA. Insert size= 1107 bp. CXCR30TN00 Chemokine (C-X-C motif) receptor 3 (CXCR3) CMKL100000 Chemokine like receptor 1 (CMKLR1) 3xHA-tagged (N-terminus) (Alternate Symbols: DEZ, ChemR23); (a.k.a: orphan G-protein coupled receptor, Dez). GenBank ACC# AY497547: Chemokine CXCR400000 Chemokine (C-X-C motif) receptor 4 (CXCR4) like receptor 1 (CMKLR1) (wild type) cloned into pcDNA3.1+ (Alternate Symbols: HM89, LAP3, NPYR, WHIM, LESTR, (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame NPY3R, fusin, HSY3RR, NPYY3R, D2S201E); (a.k.a: was amplified by the PCR from human kidney cDNA (Clontech). Y3; chemokine (C-X-C motif), receptor The insert was sequenced. Insert size= 1116 bp. 4 (fusin). GenBank ACC# AY242129: Chemokine (C-X-C motif) receptor 4 (CXCR4) (wild type) cloned into pcDNA3.1+ CMKL200000 Chemokine like receptor 2 (CMKLR2/GPR30) (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame (Alternate Symbols: FEG-1, CMKRL2, GPCR-Br); (a.k.a: was amplified by the PCR from lung cDNA. Insert size= 1059 bp. chemokine receptor-like 2). Identical to GenBank Acc# CXCR40TN00 Chemokine (C-X-C motif) receptor 4 (CXCR4) NM_001505 with the following exception: (C906T) (Silent 3xHA-tagged (N-terminus) mutation). Chemokine like receptor 2 (CMKLR2/GPR30) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XbaI (3'). The open reading frame was amplified by PCR from human CXCR500000 Chemokine (C-X-C motif) receptor 5 (CXCR5) kidney cDNA (Clontech). Insert size= 1128 bp. (Alternate Symbols: BLR1; MDR15); (a.k.a: monocyte-derived receptor 15; C-X-C chemokine receptor type 5; Burkitt lymphoma CX3R100000 Chemokine (C-X3-C motif) receptor 1 (CX3CR1) receptor 1, GTP-binding protein). Chemokine (C-X-C motif) (Alternate Symbols: V28, CCRL1, GPR13, CMKDR1, GPRV28, receptor 5 (CXCR5) (wild type) cloned into pcDNA3.1+ CMKBRL1); (a.k.a: G protein-coupled receptor 13; chemokine (Invitrogen) at EcoRI(5') and XbaI (3'). The open reading frame (C-X3-C) receptor 1; chemokine (C-C) receptor-like 1). was amplified by the PCR from leukocyte cDNA. The insert was Chemokine (C-X3-C motif) receptor 1 (CX3CR1) (wild type) sequenced and found to be identical to GenBank ACC# cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). NM_001716 with the following exception: C498->T (silent). Insert The open reading frame was amplified by the PCR from a clone size= 1119 bp. obtained from the Laboratory of Genetics/National Institutes of CXCR50TN00 Chemokine (C-X-C motif) receptor 5 (CXCR5) Mental Health at the NIH. The insert was sequenced and found 3xHA-tagged (N-terminus) to be identical to GenBank ACC# NM_001337. Insert size= 1068 bp. CXCR600000 Chemokine (C-X-C motif) receptor 6 (CXCR6) CX3R10TN00 Chemokine (C-X3-C motif) receptor 1 (CX3CR1) (Alternate Symbols: BONZO, STRL33, TYMSTR). Identical to 3xHA-tagged (N-terminus) GenBank ACC# AY322543: Chemokine (C-X-C motif) receptor 6 (CXCR6) (wild type) cloned into pcDNA3.1+ (Invitrogen) at CXCR100000 Interleukin 8 receptor alpha (IL8RA/CXCR1) EcoRI (5') and XhoI (3'). The open reading frame was amplified (Alternate Symbols: CXCR1, IL8R1, CMKAR1, IL8RBA, by the PCR from Human Genomic DNA . The insert was CDw128a, C-C CKR-1, C-C-CKR-1); (a.k.a: IL-8 receptor; IL-8 sequenced. Insert size= 1029bp receptor type 1; chemokine (C-X-C) receptor 1; interleukin-8 CXCR60TN00 Chemokine (C-X-C motif) receptor 6 (CXCR6) receptor type 1; interleukin-8 receptor type A; high affinity 3xHA-tagged (N-terminus) interleukin-8 receptor A). Interleukin 8 receptor alpha (IL8RA/CXCR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) FKS8000000 Putative chemokine receptor (FKSG80/GPR81) at EcoRI(5') and XhoI (3'). The open reading frame was amplified (a.k.a: putative chemokine receptor T-cell activation G protein- by the PCR from a clone obtained from the Laboratory of coupled receptor). GenBank ACC# AY507143: Putative Genetics/National Institutes of Mental Health at the NIH. The chemokine receptor (FKSG80/GPR81) (wild type) cloned into insert was sequenced and found to be identical to GenBank pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open ACC# NM_000634. Insert size= 1053bp. reading frame was amplified by the PCR from human kidney CXCR10TN00 Interleukin 8 receptor alpha (IL8RA/CXCR1) cDNA (Clontech). The insert was sequenced. Insert size= 1041 3xHA-tagged (N-terminus) bp.

CXCR200000 Interleukin 8 receptor beta (IL8RB/CXCR2) XCR0100000 Chemokine (C motif) receptor 1 (XCR1) (GPR5) (Alternate Symbols: CXCR2, IL8R2, IL8RA, CMKAR2, (Alternate Symbols: GPR5, CCXCR1); (a.k.a: lymphotactin CDw128b); (a.k.a: GRO/MGSA receptor; interleukin 8 receptor receptor; chemokine (C motif) XC receptor 1). GenBank ACC# B; chemokine (CXC) receptor 2; interleukin 8 receptor type 2; AY275469: Chemokine (C motif) receptor 1 (XCR1) (GPR5) (wild CXCR2 gene for IL8 receptor type B; high affinity interleukin-8 type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI receptor B). Interleukin 8 receptor beta (IL8RB/CXCR2) (wild (3'). The open reading frame was amplified by the PCR from type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI human genomic DNA . The insert was sequenced. Insert size= (3'). The open reading frame was amplified by the PCR from a 1002 bp clone obtained from the Laboratory of Genetics/National XCR010TN00 Chemokine (C motif) receptor 1 (XCR1/GPR5) Institutes of Mental Health at the NIH. The insert was sequenced 3xHA-tagged (N-terminus) and found to be identical to GenBank ACC# M73969. Insert size= 1083 bp. CXCR20TN00 Interleukin 8 receptor beta (IL8RB/CXCR2) 3xHA- tagged (N-terminus)

12 cDNA Resource Center

Cholecystokinin Receptors (GPR77) (C5L2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was CCKAR00000 Cholecystokinin A receptor (CCKAR) amplified by the PCR from Human Genomic DNA . The insert GenBank ACC# AY322549: Cholecystokinin A receptor was sequenced. Insert size= 1014 bp (CCKAR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at C5L200TN00 G-protein-coupled receptor 77 (GPR77) (C5L2) EcoRI(5') and XhoI (3'). The open reading frame was amplified 3xHA-tagged (N-terminus) by the PCR from human stomach cDNA (Clontech). Insert size= 1287bp. C5R0100000 Complement component 5 receptor 1 (C5a ligand) CCKAR0TN00 Cholecystokinin A receptor (CCKAR) 3xHA- (C5R1) tagged (N-terminus) (Alternate Symbols: C5A, C5AR, CD88); (a.k.a: complement component-5 receptor-2 (C5a ligand). Identical to GenBank CCKBR00000 Cholecystokinin B receptor (CCKBR) ACC# NM_001736: Complement component 5 receptor 1 (C5a (Alternate Symbol: GASR); (a.k.a: CCK2 receptor; gastrin ligand) (C5R1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at receptor). GenBank ACC# AY322551: Cholecystokinin B EcoRI (5') and XhoI (3'). The open reading frame was amplified receptor (CCKBR) (wild type) cloned into pcDNA3.1+ by the PCR from Human Genomic DNA . The insert was (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame sequenced. Insert size= 1053bp was amplified by the PCR from human whole brain cDNA C5R100TN00 Complement component 5 receptor 1 (C5a ligand) (Clontech). Insert size= 1344bp. (C5R1) 3xHA-tagged (N-terminus) CCKBR0TN00 Cholecystokinin B receptor (CCKBR) 3xHA- tagged (N-terminus) Corticotropin Releasing Hormone Receptors

Coagulation Factor II Receptors CRHR100000 Corticotropin releasing hormone receptor 1 (CRHR1) F2R0000000 Coagulation factor II (thrombin) receptor (Alternate Symbols: CRF1, CRHR, CRF-R, CRFR1). GenBank (F2R/PAR1) ACC# AY457172: Corticotropin releasing hormone receptor 1 (Alternate symbols: TR, CF2R, PAR1); (a.k.a: ; (CRHR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at CC-chemokine receptor chemr1; protease-activated receptor 1). EcoRI (5') and XhoI (3'). The open reading frame was amplified Identical to GenBank Acc# BC002464: Coagulation factor II by the PCR from human brain cDNA (Clontech). The insert was (thrombin) receptor (F2R/PAR1) (wild type) cloned into sequenced. Insert size= 1248bp. pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open CRHR100000 Corticotropin releasing hormone receptor 1 reading frame was amplified by PCR from IMAGE Clone # (CRHR1) 3343051. The insert was sequenced. Insert size= 1278 bp. CRHR200000 Corticotropin releasing hormone receptor 2 F2RL100000 Coagulation factor II (thrombin) receptor-like 1 (CRHR2) (F2RL1/PAR2) (Alternate Symbols: CRF2). GenBank ACC# AY449734: (Alternate Symbols: PAR2, GPR11); (a.k.a: G protein-coupled Corticotropin releasing hormone receptor 2 (CRHR2) (wild type) receptor-11, protease-activated receptor 2). Identical to cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). GenBank Acc# BT009856 except stop codon is TGA istead of The open reading frame was amplified by the PCR from human TAG: Coagulation factor II (thrombin) receptor-like 1 brain cDNA (Clontech). The insert was sequenced. Insert size= (F2RL1/PAR2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at 1236bp. EcoRI(5') and XhoI (3'). The open reading frame was amplified CRHR200000 Corticotropin releasing hormone receptor 2 by PCR from IMAGE Clone #3857382. Insert size= 1194 bp. (CRHR2)

F2RL300000 Coagulation factor II (thrombin) receptor-like 3 (F2RL3/PAR4) Cysteinyl Leukotriene Receptors (Alternate Symbols: PAR4); (a.k.a: protease-activated receptor- 4). GenBank ACC# AY431102: Coagulation factor II (thrombin) CLR0100000 Cysteinyl 1 (CYSLTR1) receptor-like 3 (F2RL3/PAR4) (wild type) cloned into pcDNA3.1+ (Alternate Symbols: HG55, CYSLT1, CYSLTR, CYSLT1R, (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame HMTMF81, MGC46139); (a.k.a: LTD4 receptor; CysLTR vide was amplified by the PCR from human lung cDNA (Clontech). supra; cysteinyl leukotriene D4 receptor). GenBank ACC# The insert was sequenced. Insert size= 1158 bp. AY242130: Cysteinyl leukotriene receptor 1 (CYSLTR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame was amplified by the PCR from Complement Component 5a (C5a) Receptors genomic DNA. Insert size= 1014 bp. CLR010TN00 Cysteinyl leukotriene receptor 1 (CYSLTR1) C3AR100000 complement component 3a receptor 1 (C3AR1) 3xHA-tagged (N-terminus) (Alternate Symbol: AZ3B, C3AR, HNFAG09); (a.k.a: complement component 3 receptor 1). GenBank ACC# AY268431: CLR0200000 Cysteinyl leukotriene receptor 2 (CYSLTR2) complement component 3a receptor 1 (C3AR1) (wild type) (Alternate Symbols: GPCR, HG57, CYSLT2, HPN321, cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). CYSLT2R, hGPCR21, PSEC0146); (a.k.a: cysteinyl leukotriene The open reading frame was amplified by the PCR from Human CysLT2 receptor). Identical to GenBank ACC# NM_020377: Genomic DNA . The insert was sequenced. Insert size= 1449 bp Cysteinyl leukotriene receptor 2 (CYSLTR2) (wild type) cloned C3AR10TN00 Complement component 3a receptor 1 (C3AR1) into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open 3xHA-tagged (N-terminus) reading frame was amplified by the PCR from genomic DNA. Insert size= 1041 bp. C5L2000000 G-protein-coupled receptor (C5L2) CLR020TN00 Cysteinyl leukotriene receptor 2 (CYSLTR2) (Alternate Symbol: GPR77); (a.k.a: G protein-coupled receptor 3xHA-tagged (N-terminus) 77). GenBank ACC# AY268430: G-protein-coupled receptor 77

13 cDNA Resource Center

EDNRB00000 type B (EDNRB) Dopamine Receptors (Alternate Symbols: ETB, ETRB, HSCR, ABCDS, HSCR2) (a.k.a: Hirschsprung disease 2). GenBank ACC# AY275463: DRD0100000 D1 (DRD1) Endothelin receptor type B (EDNRB) (wild type) cloned into (Alternate Symbols: DADR, DRD1A). GenBank Accession # pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open AF498961: Human (wild type) cloned into reading frame was amplified by the PCR from human placenta pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open cDNA . The insert was sequenced. Insert size= 1329 bp reading frame was amplified by the PCR from human whole EDNRB0TN00 Endothelin receptor type B (EDNRB) 3xHA- brain cDNA (Clontech). The insert was sequenced. Insert size = tagged (N-terminus) 1340 bp. DRD010TN00 Dopamine receptor D1 (DRD1) 3xHA-tagged (N- terminus) Estrogen Receptor

DRD0200000 Dopamine receptor 2, transcript variant 1 (DRD2) GPER100000 Estrogen receptor 1 (GPER1) (Alternate Symbols: D2R, D2DR). Identical to GenBank (a.k.a: chemokine receptor-like 2,CMKLR2). Identical to ACC#NM_000795: Dopamine receptor 2, transcript variant 1 GenBank Acc# NM_001505 with the following exception: (DRD2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at (C906T) (Silent mutation). Human Estrogen receptor 1 (GPER1) EcoRI(5') and XhoI (3'). The open reading frame was amplified (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and by the PCR from IMAGE clone ID# 3939741. Insert size= 1332 XbaI (3'). The open reading frame was amplified by PCR from bp. human kidney cDNA (Clontech). The insert was sequenced. DRD020TN00 (DRD2) 3xHA-tagged (N- Insert size = 1128 bp. *Note GPER1 was once sold as CMKL2. terminus) GPER10TN00 Estrogen receptor 1 (GPER1) 3xHA-tagged (N- terminus) DRD03A0000 (DRD3), transcript variant a (Alternate Symbols: DRD3a, D3DR, EMT1, FET1). GenBank Follicle Stimulating Receptor ACC# NM_00796: Dopamine Receptor D3a (DRD3a) (wild type) cloned into pcDNA3.1+ (Invitrogen) at HindIII(5') and XhoI(3'). FSHR000000 Follicle stimulating hormone receptor (FSHR) The open reading frame was amplified by the PCR from whole (Alternate Symbols: ODG1, FSHRO); (a.k.a: FSH receptor, brain cDNA. Insert size= 1203 bp. follitropin receptor, ovarian dysgenesis 1). GenBank ACC# AY429104: Follicle stimulating hormone receptor (FSHR) (wild DRD0400000 (DRD4) (Alternate type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI Symbols: D4DR). GenBank ACC# NM_00797: Dopamine (3'). The open reading frame was amplified by the PCR from Receptor D4 (DRD4) (wild type) cloned into pcDNA3.1+ Human testis cDNA (Clontech) . The insert was sequenced. (Invitrogen) at KpnI(5') and EcoRI(3'). The open reading frame Insert size= 2088bp was amplified by the PCR from whole brain cDNA. Insert size= 1260 bp. Formyl Peptide Receptors DRD0500000 (DRD5)

(Alternate Symbols: DBDR, DRD1B, DRD1L2, MGC10601); FPR0100000 1 (FPR1) (a.k.a: dopamine receptor D1B; D1beta dopamine receptor). Identical to GenBank ACC# NM_002029: Formyl peptide GenBank Accession # AY136750: Human dopamine receptor D5 receptor 1 (FPR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and at EcoRI (5') and XhoI (3'). The open reading frame was XhoI (3'). The open reading frame was amplified by the PCR amplified by the PCR from Human Genomic DNA . The insert from IMAGE clone (3928370). The insert was sequenced. Insert was sequenced. Insert size= 1053 bp size = 1430 bp. FPR010TN00 Formyl peptide receptor 1 (FPR1) 3xHA-tagged DRD050TN00 Dopamine receptor D5 (DRD5) 3xHA-tagged (N- (N-terminus) terminus)

FPRL100000 Formyl peptide receptor-like 1 (FPRL1)

GenBank ACC# AY225226: formyl peptide receptor-like 1 Endothelin Receptors (FPRL1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the EDNBL00000 Endothelin receptor type B-like (EDNRBL) PCR from Human Genomic DNA . The insert was sequenced. (Alternate symbols: PAELR; GPR37) (a.k.a: hET(B)R-LP; Parkin- Insert size= 1056 bp associated endothelin receptor-like receptor). Identical to FPRL10TN00 Formyl peptide receptor-like 1 (FPRL1) 3xHA- GenBank Acc# NM_005302: Endothelin Receptor type B-like tagged (N-terminus) (EDNRBL) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame was amplified FPRL200000 Formyl peptide receptor-like 2 (FPRL2) by PCR from IMAGE clone (ID# 5174687). Insert size= 1842 bp. GenBank ACC# AY262690: Formyl peptide receptor like 2 (FPRL2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI EDNRA00000 Endothelin receptor type A (EDNRA) (5') and XhoI (3'). The open reading frame was amplified by the (Alternate Symbols: ETA, ETRA). GenBank ACC# AY275462: PCR from Human Genomic DNA . The insert was sequenced. Endothelin receptor type A (EDNRA) (wild type) cloned into Insert size= 1062bp pcDNA3.1+ (Invitrogen) at HindIII (5') and XhoI (3'). The open FPRL20TN00 Formyl peptide receptor-like 2 (FPRL2) 3xHA- reading frame was amplified by the PCR from human placenta tagged (N-terminus) cDNA . The insert was sequenced. Insert size= 1284 bp EDNRA0TN00 Endothelin receptor type A (EDNRA) 3xHA- tagged (N-terminus)

14 cDNA Resource Center

Free Fatty Acid Receptors mutation). (GCGR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open FFAR100000 1 (FFAR1) reading frame was amplified by PCR from human liver cDNA (Alternate Names: FFA1R, GPR40). Accession number: (Clontech). Insert size= 1434 bp. NM_005303. Human free fatty acid receptor 1 (FFAR1), wild type, cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and GLP2R00000 Glucagon-like peptide 2 receptor (GLP2R) XhoI(3'). Contains one non-silent mutation at nt632 (G->A) Identical to GenBank Acc# NM_004246 with the following resulting in an change of Arg to His. The open reading frame exceptions: (C510T, A1326G) (both silent mutations). Glucagon- was amplified by PCR from human genomic DNA. Insert size= like peptide 2 receptor (GLP2R) (wild type) cloned into 903 bp. pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame was amplified by PCR from human stomach cDNA FFAR200000 Free Fatty Acid Receptor 2 (FFAR2) (Clontech). Insert size= 1662 bp. (Alternate Names: FFA2R, GPR43). Accession number: NM_005306. Human free fatty acid receptor 2 (FFAR2), wild type, cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and Glutamate Metabotropic Receptors XhoI(3'). The open reading frame was amplified by PCR from human genomic DNA. Insert size= 993 bp. GRM2000000 Human , Metabotropic 2 (GRM2) FFAR300000 Free Fatty Acid Receptor 3 (FFAR3) (Alternate Symbols: GLUR2, mGlu2, GPRC1B, MGLUR2). (Alternate Names: FFA3R, GPR41). Accession number: GenBank ACC# NM_000839: Human Glutamate Receptor, NM_005304. Human free fatty acid receptor 3 (FFAR3), wild Metabotropic 2 (GRM2) (wild type) cloned into pcDNA3.1+ type, cloned into pcDNA3.1+ (Invitrogen) at HindIII(5') and (Invitrogen) at BamHI(5') and XhoI(3'). The open reading frame XhoI(3'). There is one non-silent mutation at nt91 C->G resulting was amplified by PCR from whole-brain cDNA. Insert size= 2619 in a Lue to Val substitution. The open reading frame was bp. amplified by PCR from pituitary cDNA. Insert size= 1041 bp. GRM3000000 Human Glutamate Receptor, Metabotropic 3 (GRM3) Frizzled Homolog Receptor (Alternate Symbols: GLUR3, mGlu3, GPRC1C, MGLUR3). GenBank ACC# NM_000840: Human Glutamate Receptor, FZD4000000 Frizzled homolog 4 (Drosophila) (FZD4) Metabotropic 3 (GRM3) (wild type) cloned into pcDNA3.1+ (Alternate Symbols: EVR1, Fz-4, FzE4, GPCR, FZD4S, (Invitrogen) at EcoRI(5') and XhoI(3'). The open reading frame MGC34390); (a.k.a: frizzled 4, WNT receptor frizzled-4). was amplified by PCR from whole-brain cDNA. Insert size= 2640 GenBank ACC# AY462097: Frizzled homolog 4 (Drosophila) bp. (FZD4) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the GRM8000000 Human Glutamate Receptor, Metabotropic 8 PCR from Human heart cDNA (Clontech) . The insert was (GRM8), wild-type sequenced. Insert size= 1614bp (Alternate Names: GLUR8, mGlu8, GPRC1H). Accession number: NM_000845. Human glutamate receptor, Metabotropic 8 (GRM8), wild type, cloned into pcDNA3.1+ (Invitrogen) at Galanin Receptors HindIII(5') and XhoI(3'). This version of GRM8 has been mutated from catalog# GRM800000M to match the concensus sequence.

Insert size= 2727 bp. GALR100000 1 (GALR1)

(Alternate Symbols: GALNR, GALNR1); (a.k.a: Galanin GRM800000M Human Glutamate Receptor, Metabotropic 8 receptor). GenBank ACC# AY541036: (GRM8), mutant (GALR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (Alternate Symbols: GLUR3, mGlu3, GPRC1C, MGLUR3). (5') and XhoI (3'). The open reading frame was amplified by the GenBank ACC# NM_000845: Human Glutamate Receptor, PCR from human brain cDNA (Clontech). The insert was Metabotropic 8 (GRM8) (wild type) cloned into pcDNA3.1+ sequenced. Insert size= 1050 bp. (Invitrogen) at HindIII(5') and XbaI(3'). Matches above accession

number with the following non-silent mutations; nt1772 C->T (aa GALR200000 (GALR2) V->A), and nt2303 A->T (aa I->N). The open reading frame was (Alternate Symbols: GALNR2); Identical to GenBank ACC# amplified by PCR from whole-brain cDNA. Insert size= 2727 bp. NM_003857. Galanin Receptor 2 (GALR2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and EcoRI (3'). The open reading frame was amplified by the PCR from Human cDNA Gonadotropin-Releasing Hormone Receptor (hippocampus). The insert was sequenced. Insert size= 1164bp. GNRHR00000 Gonadotropin-releasing hormone receptor GALR300000 (GALR3) (GNRHR) GenBank ACC# AY587582: Galanin receptor 3 (GALR3) (wild (Alternate Symbol: GRHR, LHRHR); (a.k.a: gonadotropin- type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI releasing hormone receptor (leutinizing-releasing hormone (3'). The open reading frame was amplified by the PCR from receptor). GenBank ACC#AY39201: Gonadotropin-releasing human brain cDNA (Clontech). The insert was sequenced. Insert hormone receptor (GNRHR) (wild type) cloned into pcDNA3.1+ size= 1107bp. (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame was amplified by the PCR from human pituitary cDNA (Clontech). Insert size= 987 Glucagon Receptors GNRHR20000 Gonadotropin-releasing hormone receptor varient

2, (GNRHR2) GCGR000000 Glucagon receptor (GCGR) (Alternate Symbol: GRHR, LHRHR); (a.k.a: gonadotropin- (Alternate Symbols: GGR). Identical to GenBank Acc# releasing hormone receptor (leutinizing-releasing hormone NM_000160 with the following exception: (T1095C) (Silent

15 cDNA Resource Center

receptor). GenBank ACC# NM_00102763: Gonadotropin- releasing hormone receptor (GNRHR2) (wild type) cloned into Hypocretin Receptor pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame was amplified by the PCR from GNRHR00000. HCR0100000 Hypocretin () (Alternate Symbols: OX1R). Insert size= 750nt. Identical to GenBank ACC# AY062030: Hypocretin (orexin) receptor 1 (HCRTR1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame Growth Hormone Releasing Receptor was amplified by the PCR from Human brain cDNA (Clontech) . The insert was sequenced. Insert size= 1278bp GHRHR00000 Growth hormone releasing hormone receptor (GHRHR) (Alternate Symbols: GHRFR). GenBank ACC# AY557192: KISS Receptor Growth hormone releasing hormone receptor (GHRHR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI KISS1R0000 KISS receptor 1 (KISS1/GPR54 (3'). The open reading frame was amplified by the PCR from (a.k.a: GPR54). GenBank Acc# NM_032551. Human KISS human pituitary cDNA (Clontech). The insert was sequenced. receptor 1 (KISS1) (wild type) cloned into pcDNA3.1+ Insert size= 1272 bp. (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by PCR from human kidney cDNA (Clontech). The insert was sequenced. Insert size = 1197 bp. Growth Hormone Secretagogue Receptors

GHSR0A0000 Growth hormone secretagogue receptor type 1a Leucine-Rich Repeat-Containing Receptor (GHSR1a) GenBank ACC# AY429112: Growth hormone secretagogue LGR4000000 Leucine-rich repeat-containing GPCR 4 (LGR4) receptor type 1a (GHSR1a) (wild type) cloned into pcDNA3.1+ (Alternate Symbols: GPR48). GenBank ACC# NM_018490: (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame Human Leucine-rich repeat-containing GPCR 4 (LGR4) (wild was amplified by the PCR from Human brain cDNA (Clontech) . type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and The insert was sequenced. Insert size= 1101bp XhoI(3'). Matches the above accession# with 6 nt exceptions all of which match an alternative ACC# AB_209743. Complete GHSR0B0000 Growth Hormone Secretagogue Receptor information regarding location and residue of each point mutation (GHSR1b) is in the Sequence PDF file. The open reading frame was GenBank ACC# AY322544: Growth Hormone secretagogue amplified by PCR from whole-brain cDNA. Insert size= 2856 bp. receptor (GHSR1b) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame Leokotriene B4 Receptors was amplified by the PCR from Human Genomic DNA . The insert was sequenced. Insert size= 870 bp LTB4R00000 (LTB4R)

(Alternate Symbols: BLTR, P2Y7, GPR16, LTBR1, P2RY7,

CMKRL1, LTB4R1); (a.k.a: chemokine receptor-like 1 ; G Histamine Receptors protein-coupled receptor 16 ; P2Y, G-protein coupled, 7). GenBank ACC# AY322535: leukotriene B4 receptor HRH0100000 HRH1 (LTB4R) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI GenBank Accession # AY136743: Human histamine receptor (5') and XhoI (3'). The open reading frame was amplified by the HRH1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at HindIII PCR from Human Genomic DNA . The insert was sequenced. (5') and XhoI (3'). The open reading frame was amplified by the Insert size= 1059 bp PCR from human lung cDNA (Clontech). The insert was sequenced. Insert size = 1460 bp. LTB4R20000 Leukotriene B4 receptor 2 (LTB4R2) (Alternate Symbols: BLT2, BLTR2, JULF2); (a.k.a: leukotriene HRH0200000 Histamine receptor HRH2 B4 receptor BLT2; seven transmembrane receptor BLTR2). GenBank Accession # AY136744: Human histamine receptor GenBank ACC# AY268433: leukotriene B4 receptor 2 (LTB4R2) HRH2 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (5') and XhoI (3'). The open reading frame was amplified by the XhoI (3'). The open reading frame was amplified by the PCR PCR from human stomach cDNA (Clontech). The insert was from Human Genomic DNA . The insert was sequenced. Insert sequenced. Insert size = 1080 bp. size= 1170 bp HRH020TN00 Histamine receptor HRH2 3xHA-tagged (N- terminus) Lysophosphatidic Acid/S1P Receptors HRH0300000 Histamine receptor 3 (HRH3)

(Alternate Names: HH3R, GPCR97). Accession number: EDG0100000 Sphingosine 1-phosphate receptor 1 NM_007232. Human histamine receptor 3 (HRH3), wild type, (S1P1/EDG1) cloned into pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI(3'). (Alternate Symbols: ECGF1, CHEDG1, D1S3362); (a.k.a: edg- This construct was donated to the collection from an external lab 1; sphingosine 1- phosphate receptor EDG1; G protein- coupled and mutated to match consensus. Insert size= 1340 bp. sphingolipid receptor). Identical to GenBank ACC# NM_001400:

with the following exeption (T558C) (Silent mutation) Endothelial HRH0400000 Histamine receptor HRH4 differentiation G- protein coupled receptor 1 (EDG1) (wild type) GenBank Accession # AY136745: Human histamine receptor cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). HRH4 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI The open reading frame was amplified by the PCR from Human (5') and XhoI (3'). The open reading frame was amplified by the Genomic DNA . The insert was sequenced. Insert size= 1149 bp PCR from human leukocyte cDNA (Clontech). The insert was EDG010TN00 Sphingosine 1-phosphate receptor 1 sequenced. Insert size = 1180 bp. (S1P1/EDG1) 3xHA-tagged (N-terminus)

16 cDNA Resource Center

testis cDNA (Clontech) . The insert was sequenced. Insert size= EDG0200000 Lysophosphatidic acid receptor 1 (LPA1/EDG2) 1062 bp (Alternate Symbols: LPA1, edg-2, vzg-1, Gpcr26, Mrec1.3, EDG070TN00 Lysophosphatidic acid receptor 3 (LPA3/EDG7) rec.1.3); (a.k.a: ventricular zone gene 1). GenBank ACC# 3xHA-tagged (N-terminus) AY322546: Endothelial differentiation G-protein coupled receptor 2 (EDG2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EDG0800000 Sphingosine 1-phosphate receptor 5 BamHI (5') and XhoI (3'). The open reading frame was amplified (S1P5/EDG8) by the PCR from whole brain cDNA (Clontech) . The insert was (Alternate Symbols: S1P5, Edg-8, SPPR- 1, SPPR-2); (a.k.a: sequenced. Insert size= 1095 bp sphingosine 1- phosphate receptor 5; sphingosine 1- phosphate EDG020TN00 Lysophosphatidic acid receptor 1 (LPA1/EDG2) receptor Edg-8). GenBank ACC# AY262689: Endothelial 3xHA-tagged differentiation, sphingolipid G-protein- coupled receptor, 8 (EDG8) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI EDG0300000 Sphingosine 1-phosphate receptor 3 (5') and XhoI (3'). The open reading frame was amplified by the (S1P3/EDG3) PCR from Human Genomic DNA . The insert was sequenced. (Alternate Symbols: LPB3, S1P3, EDG- 3); (a.k.a: S1P receptor Insert size= 1197 bp EDG3; sphingosine 1-phosphate receptor 3; G protein-coupled EDG080TN00 Sphingosine 1-phosphate receptor 5 receptor endothelial differentiation gene-3). Human Endothelial (S1P5/EDG8) 3xHA-tagged (N-terminus) differentiation, sphingolipid G-protein-coupled receptor, 3 (EDG3) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the MAS-Related Receptors PCR from genomic DNA. The insert was sequenced and found to be identical with GB ACC# NM_005226 with the following MRGX100000 MAS-related GPR, member X1 (MRGPRX1) exception: G962->A (R321- >Q). Insert size = 1137 bp. (Alternate Names: MRGX1). Accession number: NM_147199. EDG030TN00 Sphingosine 1-phosphate receptor 3 Human MAS-related GPR, member X1 (MRGPRX1) (wild type) (S1P3/EDG3) 3xHA-tagged (N-terminus) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). The open reading frame was amplified by PCR from human EDG0400000 Lysophosphatidic acid receptor 2 (LPA2/EDG4) genomic DNA. Construct contains two mutations; A138G (silent) (Alternate Symbols: LPA2, EDG-4); (a.k.a: LPA receptor EDG4; and G963C resulting in residue E321D amino acid substitution. G protein- coupled receptor Lysophosphatidic acid receptor The insert was sequenced. Insert size = 969 bp. EDG4). GenBank ACC# AY322548: Endothelial differentiation G- protein coupled receptor 4 (EDG4) (wild type) cloned into MRGX200000 MAS-related GPR, member X2 (MRGPRX2) pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open (Alternate Names: MRGX2). Accession number: NM_054030. reading frame was amplified by the PCR from human MCF-7 Human MAS-related GPR, member X2 (MRGPRX2) (wild type) breast cancer cell line cDNA (in house) . The insert was cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). sequenced. Insert size= 1056 bp The open reading frame was amplified by PCR from human EDG040TN00 Lysophosphatidic acid receptor 2 (LPA2/EDG4) genomic DNA. Construct contains one mutation: A185G resulting (Alternate Symbols: LPA2, EDG-4); 3xHA-tagged (N-terminus) in residue N62S amino acid substitution. The insert was sequenced. Insert size = 993 bp. EDG0500000 Sphingosine 1-phosphate receptor 2 (S1P2/EDG5) MRGX300000 MAS-related GPR, member X3 (MRGPRX3) (Alternate Symbols: H218, LPB2, S1P2, AGR16, EDG-5, (Alternate Names: MRGX3). Accession number: AY042215. Gpcr13); (a.k.a: S1P receptor EDG5; sphingosine 1- phosphate Human MAS-related GPR, member X3 (MRGPRX3) (wild type) receptor 2). GenBank ACC# AY262688: Endothelial cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). differentiation, sphingolipid G-protein-coupled receptor, 5 Matches above accession number perfectly but contains 3nt (EDG5) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI differences resulting in 2aa residue changes from (5') and XbaI (3'). The open reading frame was amplified by the acc#NM_054031. The open reading frame was amplified by PCR from Human Genomic DNA . The insert was sequenced. PCR from whole-brain cDNA. Insert size= 969 bp. Insert size= 1062 bp EDG050TN00 Sphingosine 1-phosphate receptor 2 Melanin-Concentrating Hormone Receptors (S1P2/EDG5) 3xHA-tagged (N-terminus)

MCHR100000 Melanin-concentrating hormone receptor 1 EDG0600000 Sphingosine 1-phosphate receptor 4 (MCHR1/GPR24) (S1P4/EDG6) (Alternate Symbols: SLC1, GPR24, MGC32129); (a.k.a: G (Alternative Symbols: LPC1, SLP4); GenBank ACC# AY322537: protein-coupled receptor 24). GenBank ACC# AY562945: Endothelial differentiation G-protein-coupled receptor, 6 (EDG6) Melanin-concentrating hormone receptor 1 (MCHR1/GPR24) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR XhoI (3'). The open reading frame was amplified by the PCR from Human Genomic DNA . The insert was sequenced. Insert from human brain cDNA (Clontech). The insert was sequenced. size= 1155 bp Insert size= 1269 bp. EDG060TN00 Sphingosine 1-phosphate receptor 4

(S1P4/EDG6) 3xHA-tagged (N-terminus) MCHR200000 Melanin-concentrating hormone receptor 2 (MCHR2/GPR145) EDG0700000 Lysophosphatidic acid receptor 3 (LPA3/EDG7) (Alternate Symbols: SLT, MCH2, GPR145, MCHR2); (a.k.a: G (Alternative Symbols: GPCR, Edg-7, LP- A3, HOFNH30, RP4- protein-coupled receptor slt, G protein-coupled receptor 145). 678I3); (a.k.a: LPA receptor EDG7 ; endothelial cell GenBank ACC# AY562946: Melanin-concentrating hormone differentiation gene 7; calcium-mobilizing lysophosphatidic acid receptor 2 (MCHR2/GPR145) (wild type) cloned into pcDNA3.1+ receptor LP-A3). GenBank ACC# AY322547: Endothelial (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame differentiation G-protein coupled receptor 7 (EDG7) (wild type) was amplified by the PCR from human brain cDNA (Clontech). cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The insert was sequenced. Insert size= 1023 bp. The open reading frame was amplified by the PCR from human

17 cDNA Resource Center

Melanocortin Receptors MTNR1L0000 Melatonin-related receptor (MTR1L MCR0100000 (MC1R) alpha (Alternate Names: MTNR1L, GPR50). Accession number: melanocyte stimulating hormone receptor NM_004224. Human melatonin-related receptor (MTR1L), wild Identical to GenBank ACC# NM_002386: Melanocortin 1 type, cloned into pcDNA3.1+ (Invitrogen) at BamHI(5') and receptor (MC1R) alpha melanocyte stimulating hormone receptor XhoI(3'). This sequence differs from consensus above due to the (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and following (non-silent) mutations: nt958 C->T, nt1238 G->C, and XhoI (3'). The open reading frame was amplified by the PCR nt1804 G->A. The open reading frame was amplified by PCR from Human Genomic DNA . The insert was sequenced. Insert from pituitary cDNA. Insert size= 1842 bp. size= 954bp MCR010TN00 Melanocortin 1 receptor (MC1R) 3xHA-tagged (N- terminus)

MTLR100000 Motilin recepto MCR0200000 Melanocortin 2 receptor (adrenocorticotropic Motilin receptor (MTLR1) (wild type) cloned into pcDNA3.1+ hormone receptor): MC2R/ACTHR (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame Identical to GenBank ACC# NM_000529: Melanocortin 2 was amplified by the PCR from human stomach cDNA receptor hormone receptor (adrenocorticotropic hormone (Clontech). The insert was sequenced. Insert size= 1239 bp receptor): MC2R/ACTHR (wild type) cloned into pcDNA3.1+

(Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR from Human Genomic DNA . The insert was sequenced. Insert size= 894bp Neuromedin U Receptors MCR020TN00 Melanocortin 2 receptor (MC2R) 3xHA-tagged (N- terminus) NMU1R00000 1R (NMU1R) (Alternate Symbols: GPR66, FM-3). Identical to GenBank Acc# MCR0300000 (MC3R) NM_006056: Neuromedin U receptor 1R (NMU1R) (wild type) GenBank ACC# AY227893: melanocortin 3 receptor MC3R (wild cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI The open reading frame was amplified by PCR from IMAGE (3'). The open reading frame was amplified by the PCR from Clone #6194542. Insert size= 1281 bp. Human Genomic DNA . The insert was sequenced. Insert size= NMU1R0TN00 (NMU1R) 3xHA-tagged 1083 bp (N-terminus) MCR030TN00 Melanocortin 3 receptor (MC3R) 3xHA-tagged (N- terminus) NMU2R00000 Neuromedin U receptor 2R (NMU2R) (Alternate Symbol: FM4). Identical to GenBank Acc# NM_020167: Neuromedin U receptor 2R (NMU2R) (wild type) MCR0400000 (MC4R) cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). GenBank ACC# AY236539: melanocortin 4 receptor MC4R (wild The open reading frame was amplified by PCR from IMAGE type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI Clone #3852151. Insert size= 1248 bp. (3'). The open reading frame was amplified by the PCR from NMU2R0TN00 (NMU2R) 3xHA-tagged Human Genomic DNA . The insert was sequenced. Insert size= (N-terminus) 999 bp

MCR040TN00 Melanocortin 4 receptor (MC4R) 3xHA-tagged (N- Neuropeptide FF Receptor terminus)

NPFFR10000 Neuropeptide FF receptor 1 (NPFFR1) MCR0500000 (MC5R) (Alternate Names: NPFF1, GPR147). Accession number: Identical to GenBank ACC# NM_005913: melanocortin 5 NM_022146. Human neuropeptide FF receptor 1 (NPFFR1), wild receptor MC5R (wild type) cloned into pcDNA3.1+ (Invitrogen) at type, cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and BamHI (5') and XhoI (3'). The open reading frame was amplified XhoI(3'). The open reading frame was amplified by PCR from by the PCR from Human Genomic DNA . The insert was spinal cord cDNA. Insert size= 1293 bp. sequenced. Insert size= 978 bp

MCR050TN00 Melanocortin 5 receptor (MC5R) 3xHA-tagged (N- terminus) Neuropeptide Y Receptors

NPYR100000 Neuropeptide Y receptor Y1 (NPY1R) (Alternate Symbol: NPYR); (a.k.a: Neuropeptide Y receptor). GenBank ACC# AY548168: neuropeptide Y receptor Y1

(NPY1R) (wild type) cloned into pcDNA3.1+ (Invitrogen) at MTNR1B0000 (MTNR1B) BamHI (5') and XbaI (3'). The open reading frame was amplified (Alternate Symbols: MEL-1B-R); (a.k.a: melatonin receptor type by the PCR from human whole brain cDNA (clontech). The insert 1B). GenBank ACC# AY521019: Melatonin receptor 1B was sequenced. Insert size= 1155 bp (MTNR1B) (wild type) cloned into pcDNA3.1+ (Invitrogen) at NPYR10TN00 Neuropeptide Y1 Receptor (NPY1R) 3xHA tagged EcoRI (5') and XbaI (3'). The open reading frame was amplified (N-terminus) by the PCR from human retina cDNA (Clontech). The insert was sequenced. Insert size= 1089 bp. NPYR200000 neuropeptide Y receptor Y2 (NPY2R)

GenBank ACC# AY236540: neuropeptide Y receptor Y2 MTNR1B0000 Melatonin receptor 1B (NPY2R) (wild type) cloned into pcDNA3.1+ (Invitrogen) at Melatonin receptor 1B (MTNR1B) (wild type) cloned into EcoRI (5') and XhoI (3'). The open reading frame was amplified pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open by the PCR from Human Genomic DNA . The insert was reading frame was amplified by the PCR from human retina sequenced. Insert size= 1146 bp cDNA (Clontech). The insert was sequenced. Insert size= 1089 NPYR20TN00 Neuropeptide Y2 Receptor (NPY2R) 3xHA tagged bp. (N-terminus)

18 cDNA Resource Center

NPYR400000 neuropeptide Y receptor Y4 (NPY4R) cDNA (whole brain). The insert was sequenced. Insert size= (Alternate Symbols: Y4, PP1, PPYR1); (a.k.a: pancreatic 1002 bp. polypeptide receptor 1). GenBank ACC# AY268432: neuropeptide Y receptor Y4 (NPY4R) (wild type) cloned into P2RY140000 Purinergic Receptor 14 (P2RY14), transcript pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open variant 2 reading frame was amplified by the PCR from Human Genomic (Alternate Symbols: P2Y14, GPR105). GenBank ACC# DNA . The insert was sequenced. Insert size= 1128 bp NM_014879: Purinergic receptor 14 (P2RY14) (wild type) cloned NPYR40TN00 Neuropeptide Y4 Receptor (NPY4R) 3xHA tagged into pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI(3'). (N-terminus) Sequence matches above acc# with one exception: nt160 G->A resulting in a Gly to Lys AA residue change. The open reading NPYR500000 neuropeptide Y receptor Y5 (NPY5R) frame was amplified by the PCR from heart cDNA. Insert size= (Alternate Symbols: NPYR5). GenBank ACC# AY322538: 1017 bp. neuropeptide Y receptor Y5 (NPY5R) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open P2RY400000 Purinergic receptor 4 (P2RY4) reading frame was amplified by the PCR from Human Genomic (a.k.a: P2Y purinoceptor 4; uridine nucleotide receptor). Identical DNA . The insert was sequenced. Insert size= 1338 bp to GenBank ACC# NM_002565: Purinergic receptor P2RY NPYR50TN00 Neuropeptide Y5 Receptor (NPY5R) 3xHA tagged (P2Y4) (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (N-terminus) (5') and XhoI (3'). The open reading frame was amplified by the PCR from Human Genomic DNA. The insert was sequenced. Insert size= 1098 bp Neurotensin Receptors P2RY40TN00 Purinergic receptor 4 (P2RY4) 3xHA-tagged (N- terminus) NTSR100000 1 (NTSR1) (Alternate Symbols: NTR). GenBank ACC# AY429106: P2Y0100000 Purinergic receptor 1 (P2RY1) (NTSR1) (wild type) cloned into (a.k.a: ATP receptor; P2Y purinoceptor 1; platelet ADP receptor; pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open P2 purinoceptor subtype Y1). GenBank Accession # AY136752: reading frame was amplified by the PCR from Human brain Human Purinergic receptor P2RY1 (wild type) cloned into cDNA (Clontech) . The insert was sequenced. Insert size= pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open 1257bp reading frame was amplified by the PCR from human lung cDNA NTSR10TN00 Neurotensin Receptor 1 (NTSR1) 3xHA-tagged (Clontech). The insert was sequenced. Insert size = 1125 bp. (N-terminus) P2Y0200000 Purinergic receptor 2 (P2RY2) NTSR200000 (NTSR2) (a.k.a: ATP receptor; purinoceptor P2Y2; P2U purinoceptor 1; (Alternate Symbols: NTR2); (a.k.a: neurotensin receptor, type 2; P2Y purinoceptor 2; P2U nucleotide receptor). GenBank -sensitive neurotensin receptor). GenBank ACC# Accession # AY136753: Human Purinergic receptor P2RY2 (wild AY429107: Neurotensin receptor 2 (NTSR2) (wild type) cloned type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The (3'). The open reading frame was amplified by the PCR from open reading frame was amplified by the PCR from Human brain human placenta cDNA (Clontech). The insert was sequenced. cDNA (Clontech) . The insert was sequenced. Insert size= Insert size = 1135 bp. 1233bp NTSR20TN00 Neurotensin Receptor 2 (NTSR2) 3xHA-tagged P2Y0600000 Pyrimidinergic receptor 6 (P2RY6) (N-terminus) (a.k.a: P2Y6 receptor; P2 purinoceptor; P2Y purinoceptor 6; G- coupled nucleotide receptor). GenBank Accession # AF498920: Human Pyrimidinergic receptor P2RY6 (wild type) cloned into Nucleotide Receptor/P2Y pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from Image clone

(3162734). The insert was sequenced. Insert size = 995 bp. FKS7900000 Putative purinergic receptor FKSG79 P2Y060TN00 Pyrimidinergic receptor 6 (P2RY6) 3xHA-tagged Putative purinergic recptor FKSG79 (wild type) cloned into (N-terminus) pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI (3'). The open reading frame was amplified by the PCR from genomic DNA. P2Y1000000 Putative purinergic receptor 10 (P2RY10) Insert size= 1002 bp. (a.k.a: putative purinergic receptor; purinergic receptor P2Y, G- FKS790TN00 Putative purinergic receptor FKSG79 3xHA- protein coupled 10). GenBank ACC# AY275461: Purinergic tagged (N-terminus) receptor 10 (P2Y10) (wild type) cloned into pcDNA3.1+

(Invitrogen) at EcoRI (5') and Xho (3'). The open reading frame GPR3500000 G protein coupled receptor 35 (GPR35) was amplified by the PCR from Human Genomic DNA. The G protein coupled receptor 35 (GPR35) (wild type) cloned into insert was sequenced. Insert size= 1020 bp pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open P2Y100TN00 Putative purinergic receptor 10 (P2RY10) 3xHA- reading frame was amplified by the PCR from human genomic tagged (N-terminus) DNA. The insert was sequenced. Insert size= 930 bp

GPR350TN00 G protein coupled receptor 35 (GPR35) 3xHA- P2Y1100000 Purinergic receptor 11 (P2RY11) tagged (N-terminus) Purinergic receptor P2Y, G protein coupled,11 (P2RY11) (wild

type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI P2RY130000 Purinergic receptor P2Y, G-protein coupled, 13 (3'). The open reading frame was amplified by the PCR from (P2RY13) human placenta cDNA (Clontech). The insert was sequenced. (Alternate Symbols: P2Y13, GPR86,GPR94,GPCR1). Identicle Insert size= 1125 bp. to GenBank ACC# NM_023914 with the following exception: nt473T (silent). Purinergic receptor 13 (P2RY13) (wild type) P2Y1200000 Purinergic receptor 12 (P2RY12) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). (a.k.a: P2Y purinoceptor 12; ADP-glucose receptor; platelet ADP The open reading frame was amplified by the PCR from Human receptor; purinergic receptor P2RY12; Gi-coupled ADP receptor

19 cDNA Resource Center

HORK3; G-protein coupled receptor SP1999; putative G-protein coupled receptor purinergic receptor P2Y, G-protein coupled 12). GPR0010000 Orphan G Protein-Couple Receptor 1 (GPR1) GenBank Accession # AY136754: Human Purinergic receptor (Alternate Symbols: n/a). GenBank ACC# NM_005279: Orphan P2RY12 (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI G protein-coupled receptor 1 (GPR1) (wild type) cloned into (5') and XhoI (3'). The open reading frame was amplified by the pcDNA3.1+ (Invitrogen) at BamHI(5') and XhoI(3'). The open PCR from human whole brain cDNA (Clontech). The insert was reading frame was amplified by PCR from hippocampus cDNA. sequenced. Insert size = 1030 bp. Insert size= 1068 bp.

GPR0040000 Orphan G Protein-Couple Receptor 4 (GPR4) Opioid Receptors (Alternate Symbols: n/a). GenBank ACC# NM_005282: Orphan G protein-coupled receptor 4 (GPR4) (wild type) cloned into OPR0K00000 , kappa 1 (OPRK1) pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). The open (Alternate Symbols: KOR, OPRK); (a.k.a: Opiate receptor, reading frame was amplified by PCR from whole-brain cDNA. kappa-1). GenBank Accession # AF498922: Human opioid Insert size= 1089 bp receptor kappa (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was amplified by GPR0310000 G protein coupled receptor 31 (GPR31) the PCR from human whole brain cDNA (Clontech). The insert (Alternate Names: ). Accession number: NM_005299. Human was sequenced. Insert size = 1150 bp. orphan g protein coupled receptor 31 (GPR31), wild type, cloned OPR0K0TN00 Opioid Receptor Kappa (OPRK) 3xHA tagged (N- into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). There is terminus) one non-silent mutation at nt271 A->G resulting in a His to Arg substitution. The open reading frame was amplified by PCR from OPR0M10000 opioid receptor, Mu1 (OPRM1) genomic DNA. Insert size= 960 bp. (Alternate Symbols: OPRM, MOR-1). Genbank ACC# AY521028: Human opioid receptor mu1 (wild type) cloned into GPR0450000 Orphan G Protein-coupled Receptor 45 (GPR45) pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open (Alternate Symbols: PSP24, GPR45) Accession number: reading frame was amplified by the PCR from human whole NM_007227. Human orphan g protein coupled receptor 45 brain cDNA (Clontech). The insert was sequenced. Insert size = (GPR45), wild-type, cloned into pcDNA3.1+ (Invitrogen) at EcoRI 1203 bp. (5') and XhoI (3'). The open reading frame was amplified by PCR from hippocampus cDNA. Insert was sequenced. Insert = 1119. OPRD100000 Opioid receptor, delta 1 (OPRD1) (Alternate Names: OPRD, DOR-1). Accession number: GPR1320000 Orphan G Protein-Couple Receptor 132 (GPR132) NM_000911. Human opioid receptor, delta 1 (OPRD1), wild type, (Alternate Symbols: G2A). GenBank ACC# NM_013345: Orphan cloned into pcDNA3.1+ (Invitrogen) at BamHI(5') and EcoRI(3'). G protein-coupled receptor 132 (GPR132) (wild type) cloned into One silent mutation exists: nt921 C->T. The open reading frame pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). One silent was amplified by PCR from kidney cDNA. Insert size= 1119 bp. mutation at nt138 C->A but otherwise a match to the above accession number. The open reading frame was amplified by OPRL100000 Opiate receptor-like 1 (OPRL1) PCR from lung cDNA. Insert size= 1143 bp. (Alternate Symbols: OOR, ORL1, KOR-3, NOCIR); (a.k.a: ; orphanin FQ receptor; rat LC132 receptor- GPR5600000 G protein coupled receptor 56 (GPR56), variant 1 like; kappa3-related opioid receptor). GenBank ACC# (Alternate Names: GPR56 isoform a, TM7LN4). Accession AY268428: Opiate receptor-like 1 (OPRL1) (wild type) cloned number: NM_005682. Human orphan g protein coupled receptor into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The 56 (GPR56) variant 1, wild type, cloned into pcDNA3.1+ open reading frame was amplified by the PCR from Human (Invitrogen) at HindIII(5') and XbaI(3'). *** Please note, the stop Genomic DNA. The insert was sequenced. Insert size= 1113bp codon is INSIDE the XbaI(3') RE site limiting options for OPRL10TN00 Opioid Receptor-Like 1 (OPRL1) 3xHA tagged (N- subsequent sub-cloning. The open reading frame was amplified terminus) by PCR from whole brain cDNA. Insert size= 2082 bp.

GPR5700000 G protein coupled 57 (GPR57) Opsin Receptor (Alternate Symbols: n/a). G protein coupled orphan receptor 57 (GPR57) was cloned into pcDNA3.1+ (Invitrogen) at BamHI (5')

and EcoRI (3'). The open reading frame was amplified by the OPN1S00000 Opsin 1, short-wave-sensitive (OPN1SW) PCR from human genomic DNA. The insert was sequenced is (Alternative names: BCP, BOP, CBT). Protein identical to mRNA identical to GenBank Acc# AF_112461. Insert size = 1032bp. GenBank Acc# NM_001708. Human Opsin 1, short-wave- GPR570TN00 G protein coupled orphan receptor 57 (GPR57) sensitive (color-blindness, tritan) (OPN1SW) cloned into 3xHA-tagged (N-terminus) pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR from cDNA. The insert GPR6500000 Orphan G Protein-Couple Receptor 65 (GPR65) was sequenced and contains a SILENT mutation (nt366 A-C). (Alternate Symbols: n/a). GenBank ACC# NM_003608: Orphan Insert size = 1047bp. G protein-coupled receptor 65 (GPR65) (wild type) cloned into

pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). The open Orphan Receptors reading frame was amplified by PCR from thymus cDNA. Insert size= 1014 bp. ACP1R10000 Adenylate cyclase activating polypeptide 1 (pituitary) receptor type 1 (ADCYAP1R1) GPR6800000 Orphan G Protein-Couple Receptor 68 (GPR68) (Alternate symbols: PACAPR, PACAPRI). GenBank ACC# (Alternate Symbols: OGR1). GenBank ACC# NM_003485: AY366498: Adenyl cycl activating polypep 1 (pituitary) rect type 1 Orphan G protein-coupled receptor 68 (GPR68) (wild type) (ADCYAP1R1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). EcoRI(5') and XhoI (3'). The open reading frame was amplified The open reading frame was amplified by PCR from thymus by PCR from human brain cDNA (Clontech). Insert size= 1407 cDNA. Insert size= 1128 bp. Please note: this was formerly bp. found under catalog number OGR1000000.

20 cDNA Resource Center

GPR8300000 G protein coupled receptor 83 (GPR83) PTAFR00000 Platelet-activating factor receptor (PTAFR) (Alternate Names: ). Accession number: NM_016540. Human GenBank ACC# AY275466: Platelet-activating factor receptor orphan g protein coupled receptor 83 (GPR83), wild type, cloned (PTAFR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). The open (5') and XhoI (3'). The open reading frame was amplified by the reading frame was amplified by PCR from whole brain cDNA. PCR from human placenta cDNA . The insert was sequenced. Insert size= 1272 bp. Insert size= 1029 bp

GPR8700000 Orphan G Protein-Couple Receptor 87 (GPR87) (Alternate Symbols: GPR95, FKSG78, KPG_002). GenBank Prokineticin Receptors ACC# NM_023915: Orphan G protein-coupled receptor 87 (GPR87) (wild type) cloned into pcDNA3.1+ (Invitrogen) at PROKR10000 (PROKR1) HindIII(5') and XhoI(3'). One positive mutation at nt461 A->G (Alternate Symbols: GPR73, GPR73a, PKR1, ZAQ); Identical to (Serine to Asparagine) but otherwise a match to the above GenBank ACC# NM_138964. Prokineticin receptor (PROKR1) accession number. The open reading frame was amplified by (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and PCR from testis cDNA. Insert size= 1077 bp. XhoI (3'). The open reading frame was amplified by the PCR from Human cDNA (testis). The insert was sequenced. Insert GPRE200000 G-protein coupled receptor (RE2) size= 1182bp. GenBank ACC# AY275468: G-protein coupled receptor RE2 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and PROKR20000 Prokineticin Receptor (PROKR2) XhoI (3'). The open reading frame was amplified by the PCR (Alternate Symbols: GPR73b, GPR73L1, PKR2, KAL3); Identical from human brain cDNA . The insert was sequenced. Insert to GenBank ACC# NM_144773 with the following exceptions: 4 size= 1590 bp SILENT mutations (nt475T, nt595C, nt1131A, nt1119T) and 1 positive amino acid substitution at V330M. Prokineticin receptor STRG000000 G-protein coupled receptor 88 (STRG) (PROKR2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at (Alternate Symbols: GPR88). GenBank ACC# AY336999: G- BamHI (5') and EcoRI (3'). The open reading frame was protein coupled receptor 88 (STRG/GPR88) (wild type) cloned amplified by the PCR from Human cDNA (testis). The insert was into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open sequenced. Insert size= 1155bp reading frame was amplified by the PCR from Human Genomic DNA. The insert was sequenced. Insert size= 1155 bp Prolactin Releasing Hormone Receptor

Oxytocin Receptor PRLHR00000 Prolactin Releasing Hormone Receptor (PRLHR) (Alternate Symbols: GR3, GPR10). GenBank ACC# OXTR000000 (OXTR) NM_004248: human prolactin releasing hormone receptor (Alternate Symbols: OT-R). Identical to GenBank (PRLHR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at ACC#AY389507: Oxytocin receptor (OXTR) (wild type) cloned EcoRI(5') and XhoI(3'). The open reading frame was amplified by into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open the PCR from whole-brain cDNA. Insert size= 1113 bp. reading frame was amplified by the PCR from human ovary cDNA (Clontech). Insert size= 1170 bp. Prostaglandin and Thromboxane Receptors

Parathyroid Hormone Receptors CRTH200000 Prostaglandin D2 receptor, chemoattractant receptor-homologous molecule expressed on TH2 cells PTHR100000 Parathyroid hormone receptor 1 (PTHR1) (CRTH2/GPR44) (Alternate Symbols: PTHR). GenBank ACC# AY449732: (Alternate Symbol: GPR44); (a.k.a: G protein-coupled receptor Parathyroid hormone receptor 1 (PTHR1) (wild type) cloned into 44). GenBank ACC# AY507142: Prostaglandin D2 receptor, pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open chemoattractant receptor-homologous molecule expressed on reading frame was amplified by the PCR from human lung cDNA TH2 cells (CRTH2/GPR44) (wild type) cloned into pcDNA3.1+ (Clontech). The insert was sequenced. Insert size= 1782 bp. (Invitrogen) at EcoRI (5') and XbaI (3'). The open reading frame was amplified by the PCR from human placenta cDNA PTHR200000 Parathyroid hormone receptor 2 (PTHR2) (Clontech). The insert was sequenced. Insert size= 1188 bp. (a.k.a: PTH2 receptor). GenBank ACC# AY497546: Parathyroid hormone receptor 2 (PTHR2) (wild type) cloned into pcDNA3.1+ PER0100000 Prostaglandin E receptor 1, (subtype EP1), 42kDa (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame (PTGER1) was amplified by the PCR from human retina cDNA (Clontech). (Alternate Symbol: EP1); (a.k.a: Prostaglandin E receptor 1, EP1 The insert was sequenced. Insert size= 1653 bp. subtype, 42kDa). GenBank ACC# AY275470: Prostaglandin E receptor 1, subtype EP1 (PTGER1) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open Platelet Activating Factor Receptors reading frame was amplified by the PCR from human kidney cDNA. The insert was sequenced. Insert size= 1209 bp HUMNP00000 Leukocyte Platelet-Activating Factor Receptor (HUMNPIIY20) PER0200000 Prostaglandin E receptor 2, (subtype EP2), 53kDa (Alternate symbols: PAFR, GPR135I); (a.k.a: G protein-coupled (PTGER2) receptor 135; platelet activating factor receptor). Identical to (Alternate Symbol: EP2); (a.k.a: Prostaglandin E receptor 2, EP2 GenBank Acc# NM_022571: Leukocyte Platelet-Activating subtype, 53kD). GenBank ACC# AY275471: Prostaglandin E Factor Recpt (HUMNPIIY20) (wild type) cloned into pcDNA3.1+ receptor 2, subtype EP2 (PTGER2) (wild type) cloned into (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open was amplified by PCR from IMAGE clone (ID# 4825908). Insert reading frame was amplified by the PCR from human placenta size= 1485 bp. cDNA. The insert was sequenced. Insert size= 1077 bp

21 cDNA Resource Center

PER020TN00 Prostaglandin E receptor 2 (subtype EP2) XhoI (3'). The open reading frame was amplified by the PCR (PTGER2) 3xHA-tagged (N-terminus) from Human Genomic DNA. The insert was sequenced and contains one SILENT mutation nt820 T->C. This was formerly PER0400000 Prostaglandin E receptor 4 subtype EP4 sold as catalog number SALPR00000. Insert size= 1410 bp. (PTGER4) RXFP30TN00 Relaxin/Insulin-like Family Peptide Receptor 3 (Alternate Symbols: EP4, EP4R). GenBank ACC# AY429109: (RXFP3) 3xHA-tagged (N-terminus) Prostaglandin E receptor 4 subtype EP4 (PTGER4) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). RXFP400000 Relaxin/Insulin-like Family Peptide Receptor 4 The open reading frame was amplified by the PCR from Human (RXFP4) Lung cDNA (Clontech) . The insert was sequenced. Insert size= (Alternate Symbols: GPR100, RLN3R2, GPCR142). GenBank 1467bp ACC# NM_181885: Human Relaxin/Insulin-like Family Peptide Receptor 4 (RXFP4) (wild type) cloned into pcDNA3.1+ PER3VI0000 Prostaglandin E receptor 3 subtype EP3 (Isoform (Invitrogen) at BamHI(5') and EcoRI(3'). Matches above VI) (PTGER3) accession number with the following non-silent mutation; nt986 (Alternate Symbols: EP3); (a.k.a: Prostaglandin E receptor 3, T->C (aa L->S). The open reading frame was amplified by PCR EP3 subtype). GenBank ACC# AY429108: Prostaglandin E from testis cDNA. Insert size= 1125 bp. receptor 3 subtype EP3 (Isoform VI) (PTGER3) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR from Human placental cDNA (Clontech) . The insert was sequenced. Insert size= 1182bp SCTR000000 Secretin receptor (SCTR) PER3VITN00 Prostaglandin E receptor 3 (subtype EP3) (Isoform (Alternate Symbols: SR); (a.k.a: pancreatic secretin receptor). VI) (PTGER3) 3xHA-tagged (N-terminus) GenBank ACC# AY462218: Secretin receptor (SCTR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). PTGDR00000 Prostaglandin D2 Receptor (PTGDR) The open reading frame was amplified by the PCR from human (Alternate Symbols: DP, AS1, ASRT1); Identical to GenBank kidney cDNA (Clontech). The insert was sequenced. Insert size= ACC# NM_000953 with the following exceptions: nt576T 1323bp. mutation (silent). Prostaglandin D2 Receptor (PTGDR) (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and EcoRI (3'). The open reading frame was amplified by the PCR Serotonin/5-HT Receptors from Human cDNA (small intestine). The insert was sequenced.

Insert size= 1080bp. HTR01A0000 5-hydroxytryptamine (serotonin) receptor 1A (HTR1A) PTGFR00000 (FP) (PTGFR) (Alternate Symbols: ADRBRL1, ADRB2RL1). GenBank (Alternate Symbols: FP). Identical to GenBank ACC#AY337000: Accession # AF498978: Human 5-hydroxytryptamine (serotonin) Prostaglandin F receptor (FP) (PTGFR) (wild type) cloned into receptor HTR1A (wild type) cloned into pcDNA3.1+ (Invitrogen) pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open at BamHI (5') and XhoI (3'). The open reading frame was reading frame was amplified by the PCR from human placental amplified by the PCR from human whole brain cDNA (Clontech). cDNA (Clontech). Insert size= 1080 bp. The insert was sequenced. Insert size = 1270 bp. PTGFR0TN00 Prostaglandin F receptor (FP) (PTGFR) 3xHA- HTR01ATN00 5-hydroxytryptamine (serotonin) receptor 1A tagged (N-terminus) (HTR1A) 3xHA-tagged (N-terminus)

PTGIR00000 Prostaglandin I2 (prostacyclin) receptor (IP) HTR01B0000 5-hydroxytryptamine (serotonin) receptor 1B (PTGIR) (HTR1B) (Alternate Symbols: IP, PRIPR). GenBank ACC# AY242134: Alternate Symbols: S12, HTR1D2, HTR1DB); (a.k.a: 5-HT1B; 5- Prostaglandin I2 (prostacyclin) receptor (PTGIR) (wild type) HT1DB). GenBank ACC# AY225227: 5-hydroxytryptamine cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). (serotonin) receptor (HTR1B) (wild type) cloned into pcDNA3.1+ The open reading frame was amplified by the PCR from (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame placental cDNA. Insert size= 1161 bp. was amplified by the PCR from Human Genomic DNA. The PTGIR0TN00 Prostaglandin I2 (prostacyclin) receptor (PTGIR) insert was sequenced. Insert size= 1173 bp 3xHA-tagged (N-terminus) HTR01BTN00 5-hydroxytryptamine (serotonin) receptor 1B (HTR1B) 3xHA-tagged (N-terminus) TXA2R00000 Thromboxane A2 receptor (TBXA2R) GenBank ACC# AY429110: Thromboxane A2 receptor HTR01D0000 5-hydroxytryptamine (serotonin) receptor 1D (TBXA2R) (wild type) cloned into pcDNA3.1+ (Invitrogen) at (HTR1D) EcoRI (5') and XhoI (3'). The open reading frame was amplified (Alternate Symbols: HTRL; RDC4; HT1DA; HTR1DA). GenBank by the PCR from Human Placental cDNA (Clontech) . The insert Accession # AF498979: Human 5-hydroxytryptamine (serotonin) was sequenced. Insert size= 1032bp receptor HTR1D (wild type) cloned into pcDNA3.1+ (Invitrogen) TXA2R0TN00 Thromboxane A2 Receptor (TBXA2R) 3xHA- at EcoRI (5') and XbaI (3'). The open reading frame was tagged (N-terminus) amplified by the PCR from Image clone (4299633). The insert was sequenced. Insert size = 1140 bp.

Relaxin/Insulin-Like Peptide Receptors HTR01E0000 5-hydroxytryptamine (serotonin) receptor 1E (HTR1E) RXFP300000 Relaxin/Insulin-like Family Peptide Receptor 3 GenBank Accession # AF498980: Human 5-hydroxytryptamine (RXFP3) (serotonin) receptor HTR1E (wild type) cloned into pcDNA3.1+ (Alternate Symbols: SALPR). GenBank ACC# NM_016568. (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame Human Relaxin/Insulin-like Family Peptide Receptor 3 (RXFP3) was amplified by the PCR from human whole brain cDNA (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (Clontech). The insert was sequenced. Insert size = 1100 bp.

22 cDNA Resource Center

HTR01F0000 5-hydroxytryptamine (serotonin) receptor 1F GenBank ACC# AY429105: 5-hydroxytryptamine (serotonin) (HTR1F) receptor 6 (HTR6) (wild type) cloned into pcDNA3.1+ (Invitrogen) (Alternate Symbols: MR77; HTR1EL); (a.k.a: 5- at EcoRI (5') and XhoI (3'). The open reading frame was hydroxytryptamine receptor 1F). GenBank Accession # amplified by the PCR from Human brain cDNA (Clontech) . The AF498981: Human 5-hydroxytryptamine (serotonin) receptor insert was sequenced. Insert size= 1323bp HTR1F (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI HTR060TN00 5-hydroxytryptamine (serotonin) receptor 6 (5') and XhoI (3'). The open reading frame was amplified by the (HTR6) 3xHA-tagged (N-terminus) PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 1110 bp. HTR07A0000 5-hydroxytryptamine (serotonin) receptor 7A (HTR7A) HTR02A0000 5-hydroxytryptamine (serotonin) receptor 2A Identical to GenBank Acc# NM_000872: 5-hydroxytryptamine (HTR2A) (serotonin) receptor 7A (HTR7A) (wild type) cloned into (Alternate Symbol: HTR2). GenBank Accession # AF498982: pcDNA3.1+ (Invitrogen) at EcoRI(5') and Xba1 (3'). The open Human 5-hydroxytryptamine (serotonin) receptor HTR2A (wild reading frame was amplified by PCR from IMAGE Clone # type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI 5297325. The insert was sequenced. Insert size= 1338bp. (3'). The open reading frame was amplified by the PCR from HTR07ATN00 5-hydroxytryptamine (serotonin) receptor 7A human whole brain cDNA (Clontech). The insert was sequenced. (HTR7A) 3xHA-tagged (N-terminus) Insert size = 1420 bp. HTR07B0000 5-hydroxytryptamine (serotonin) receptor 7B HTR02B0000 5-hydroxytryptamine (serotonin) receptor 2B (HTR7B) (HTR2B) GenBank ACC# AY493988: 5-hydroxytryptamine (serotonin) GenBank Accession # AY136751: Human 5-hydroxytryptamine receptor 7B (HTR7B) (wild type) cloned into pcDNA3.1+ (serotonin) receptor HTR2B (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). The open reading frame (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human colon cDNA (Clontech). (Clontech). The insert was sequenced. Insert size= 1299bp. The insert was sequenced. Insert size = 1445 bp. HTR07BTN00 5-hydroxytryptamine (serotonin) receptor 7B HTR02BTN00 5-hydroxytryptamine (serotonin) receptor 2B (HTR7B) 3xHA-tagged (N-terminus) (HTR2B) 3xHA-tagged (N-terminus)

HTR02C0000 5-hydroxytryptamine (serotonin) receptor 2C Somatostatin Receptors (HTR2C) (Alternate Symbol: HTR1C). GenBank Accession # AF498983: SSTR100000 1 (SSTR1) Human 5-hydroxytryptamine (serotonin) receptor HTR2C (wild (Alternate Symbols: SRIF-2); (a.k.a: G-protein coupled receptor type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI somatostatin receptor isoform 1). GenBank ACC# AY322536: (3'). The open reading frame was amplified by the PCR from (SSTR1) (wild type) cloned into human whole brain cDNA (Clontech). The insert was sequenced. pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open Insert size = 1380 bp. reading frame was amplified by the PCR from Human Genomic HTR02CTN00 5-hydroxytryptamine (serotonin) receptor 2C DNA. The insert was sequenced. Insert size= 1176 bp (HTR2C) 3xHA-tagged (N-terminus) SSTR10TN00 Somatostatin receptor 1(SSTR1) 3xHA tagged (N- terminus) HTR03A0000 5-Hydroxytryptamine (serotonin) receptor 3A (HTR3A) SSTR200000 (SSTR2) (a.k.a: 5-HT3R; 5-hydroxytryptamine (serotonin) receptor-3). GenBank ACC# AY236542: somatostatin receptor 2 (SSTR2) GenBank Accession # AF498984: Human 5-hydroxytryptamine (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (serotonin) receptor HTR3A (wild type) cloned into pcDNA3.1+ XhoI (3'). The open reading frame was amplified by the PCR (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame from Human Genomic DNA. The insert was sequenced. Insert was amplified by the PCR from Image clone (2821710). The size= 1110 bp insert was sequenced. Insert size = 1440 bp. SSTR20TN00 Somatostatin receptor 2(SSTR2) 3xHA tagged (N- terminus) HTR0400000 5-hydroxytryptamine (serotonin) receptor 4 (HTR4) Identical to GenBank ACC# NM_000870 with the following SSTR300000 (SSTR3) exeption (C112T) silent mutation: 5-hydroxytryptamine GenBank ACC# AY322541: Somatostatin receptor 3 (SSTR3) (serotonin) receptor 4 (HTR4) (wild type) cloned into pcDNA3.1+ (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame XhoI (3'). The open reading frame was amplified by the PCR was amplified by the PCR from Human brain cDNA (Clontech) . from Human Genomic DNA. The insert was sequenced. Insert The insert was sequenced. Insert size= 1167bp size= 1257 bp HTR040TN00 5-hydroxytryptamine (serotonin) receptor 4 SSTR30TN00 Somatostatin receptor 3(SSTR3) 3xHA tagged (N- (HTR4) 3xHA-tagged (N-terminus) terminus)

HTR05A0000 5-hydroxytryptamine (serotonin) receptor 5A SSTR400000 (SSTR4) (HTR5A) GenBank ACC# AY548169: Somatostatin receptor 4 (SSTR4) GenBank Accession # AF498985: Human 5-hydroxytryptamine (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and (serotonin) receptor HTR5A (wild type) cloned into pcDNA3.1+ XhoI (3'). The open reading frame was amplified by the PCR (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame from Human Genomic DNA. The insert was sequenced. Insert was amplified by the PCR from human whole brain cDNA size= 1167bp (Clontech). The insert was sequenced. Insert size = 1070 bp. SSTR40TN00 (SSTR4) 3xHA tagged (N- HTR05ATN00 5-hydroxytryptamine (serotonin) receptor 5A terminus) (HTR5A) 3xHA-tagged (N-terminus) SSTR500000 Somatostatin receptor 5 (SSTR5) HTR0600000 5-hydroxytryptamine (serotonin) receptor 6 (HTR6) Identical to GenBank ACC# NM_001053: with the following

23 cDNA Resource Center exception (G1044A) (Silent mutation) Somatostatin receptor 5 Thyrotropin-releasing Hormone Receptor (SSTR5) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the TRHR000000 Thyrotropin-releasing hormone receptor (TRHR) PCR from Human Genomic DNA. The insert was sequenced. GenBank ACC# AY493373 : Thyrotropin-releasing hormone Insert size= 1095 bp receptor (TRHR) (wild type) cloned into pcDNA3.1+ (Invitrogen) SSTR50TN00 Somatostatin receptor 5(SSTR5) 3xHA tagged (N- at EcoRI (5') and XhoI (3'). The open reading frame was terminus) amplified by the PCR from Human pituitary cDNA (Clontech) . The insert was sequenced. Insert size= 1197bp

Succinate Receptor Trace Amine Receptors SUCNR10000 Succinate receptor 1 (SUCNR1) (Alternate Names: GPR91). Accession number: NM_033050. TAR0100000 Trace Amine Receptor (TAR) 1 Human succinate receptor 1 (SUCNR1), wild type, cloned into GenBank ACC# AY180374: Human Trace Amine Receptor pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI(3'). The open (TAR) 1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI reading frame was amplified by PCR from kidney cDNA. Insert (5') and XhoI (3'). The open reading frame was amplified by the size= 1160 bp. PCR from Human Genomic DNA. The insert was sequenced. Insert size= 1020 bp TAR010TN00 Trace Amine Receptor 1 (TAR1) 3xHA tagged (N- Tachykinin Receptors terminus)

TACR100000 1 (TACR1) TAR0300000 Trace Amine Receptor (TAR) 3 (Alternate Symbols: SPR, NK1R, TAC1R); (a.k.a: NK-1 receptor; GenBank ACC# AY183469: Human Trace Amine Receptor neurokinin 1 receptor; (substance P (TAR) 3 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI receptor; neurokinin-1 receptor); tachykinin 1 receptor (5') and XhoI (3'). The open reading frame was amplified by the (substance P receptor,neurokinin 1 receptor). GenBank ACC# PCR from Human Genomic DNA. Insert size= 1047 bp AY462098: Tachykinin receptor 1 (TACR1) (wild type) cloned TAR030TN00 Trace Amine Receptor (TAR3) 3xHA tagged (N- into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The terminus) open reading frame was amplified by the PCR from human whole brain cDNA (Clontech) . The insert was sequenced. Insert TAR0400000 Trace Amine Receptor (TAR) 4 size= 1224 bp GenBank ACC# AY183470: Human Trace Amine Receptor TACR10TN00 Tachykinin receptor 1 (TACR1) 3xHA tagged (N- (TAR) 4 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI terminus) (5') and XhoI (3'). The open reading frame was amplified by the PCR from Human Genomic DNA. Insert size= 1038 bp TACR200000 (TACR2) TAR040TN00 Trace Amine Receptor 4 (TAR4) 3xHA tagged (N- (Alternate Symbols: SKR, NK2R, NKNAR, TAC2R); (a.k.a: NK-2 terminus) receptor; neurokinin 2 receptor; Tachykinin receptor 2 (substance K receptor, neurokinin 2 receptor); tachykinin 2 TAR0500000 Trace Amine Receptor (TAR) 5 receptor (substance K receptor, neurokinin 2 receptor). GenBank GenBank ACC# AY183468: Human Trace Amine Receptor ACC# AY322545: Tachykinin receptor 2 (TACR2) (wild type) (TAR) 5 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XbaI (3'). (5') and XhoI (3'). The open reading frame was amplified by the The open reading frame was amplified by the PCR from human PCR from Human Genomic DNA. Insert size=1038 bp stomach cDNA (Clontech). The insert was sequenced. Insert TAR050TN00 Trace Amine Receptor 5 (TAR5) 3xHA tagged (N- size= 1197 bp terminus) TACR20TN00 Tachykinin receptor 2 (TACR2) 3xHA tagged (N- terminus) Transmembrane 7 Superfamily receptor TACR300000 (TACR3) (Alternate Symbols: NK3R, TAC3RL); (a.k.a: NK-3 receptor; TM7S100000 Similar to transmembrane 7 superfamily neurokinin B receptor). GenBank ACC# AY462099: Tachykinin member 1 (TM7SF1) receptor 3 (TACR3) (wild type) cloned into pcDNA3.1+ GenBank ACC# AY242135: Similar to transmembrane 7 (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame superfamily member 1 (TM7SF1) (wild type) cloned into was amplified by the PCR from human whole brain cDNA pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). The open (Clontech) The insert was sequenced. Insert size= 1398 bp reading frame was amplified by the PCR from kidney cDNA. TACR30TN00 Tachykinin receptor 3 (TACR3) 3xHA tagged (N- Insert size= 1200 bp terminus)

Urotensin Receptor Thyroid Stimulating Hormone Receptor UTR0200000 Urotensin-II receptor (UTR2/GPR14) TSHR000000 Thyroid stimulating hormone receptor (TSHR) (Alternate Symbols: UTR, GPR14); (a.k.a: G protein-coupled (A.k.a: ). GenBank ACC# AY429111: Thyroid receptor 14). Identical to GenBank Acc# NM_018949 with the stimulating hormone receptor (TSHR) (wild type) cloned into following exception: (C51T) (Silent mutation). Urotensin-II pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open receptor (UTR2/GPR14) (wild type) cloned into pcDNA3.1+ reading frame was amplified by the PCR from Human thyroid (Invitrogen) at EcoRI(5') and XhoI (3'). The open reading frame cDNA (Clontech). The insert was sequenced. Insert size= was amplified by PCR from human heart cDNA (Clontech). Insert 2295bp size= 1170 bp.

24 cDNA Resource Center

Vasoactive Intestinal Peptide Receptor VIPR200000 Vasoactive intestinal peptide receptor 2 (VIPR2) Identical to GenBank ACC# NM_003382 with the following VIPR100000 Vasoactive intestinal peptide receptor 1 (VIPR1) exception: (C825T) (Silent mutation). Vasoactive Intestinal Pep (Alternate Symbols: n/a) Identical to GenBank ACC# Rec 2 (VIPR2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at NM_004624 with the following exception: (T444C-Silent EcoRI(5') and XhoI (3'). The open reading frame was amplified mutation). Vasoactive Intestinal Pep Rec 1 (VIPR1) (wild type) by PCR from human placenta cDNA (Clontech). Insert size= cloned into pcDNA3.1+ (Invitrogen) at EcoRI(5') and XhoI (3'). 1317 bp. The open reading frame was amplified by PCR from human lung cDNA (BioChain). Insert size= 1374 bp.

25 cDNA Resource Center

GPCR Regulators

Arrestins GRB Protein RAMP Proteins

Arrestins

ARR3000000 3, retinal (X-arrestin) (ARR3) RAMP200000 Receptor (calcitonin) activity modifying protein 2 (Alternate Symbols: ) Accession number: NM_004312. Human (RAMP2) arrestin 3, retinal (X-arrestin) (ARR3), wild-type, cloned into (a.k.a: receptor-activity-modifying protein 2 calcitonin receptor- pcDNA3.1+ (Invitrogen) at HindIII (5') and XhoI (3'). The open like receptor activity modifying protein 2). GeneBank Accession # reading frame was amplified by PCR from whole-eye cDNA. AY265458. RAMPs are type I transmembrane proteins with an Insert was sequenced. Insert size = 1 extracellular N terminus and a cytoplasmic C terminus. RAMPs are required to transport calcitonin-receptor-like receptor (CRLR) ARRB200000 Arrestin, beta 2 (ARRB2), transcript variant 1 to the plasma membrane. CRLR, a receptor with seven (Alternate Symbols: ) Accession number: NM_004313. Human transmembrane domains, can function as either a calcitonin- arrestin, beta 2 (ARRB2), transcript variant 1, wild-type, cloned gene-related peptide (CGRP) receptor or an adrenomedullin into pcDNA3.1+ (Invitrogen) at BamHI (5') and EcoRI (3'). The receptor, depending on which members of the RAMP family are open reading frame was amplified by PCR from lung cDNA. expressed. In the presence of this (RAMP2) protein, CRLR Insert was sequenced. Insert size = 1230. functions as an adrenomedullin receptor. The RAMP2 protein is involved in core glycosylation and transportation of adrenomedullin receptor to the cell surface. Receptor (calcitonin) GRB Protein activity modifying protein 2 (RAMP2) (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open

reading frame was amplified by the PCR from human Heart GRB0200000 GRB2 cDNA (Clontech). The insert was sequenced. Insert size= 528 Human -bound protein 2 (GRB2) (wild RAMP20TN00 Receptor (calcitonin) activity modifying protein 2 type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho (RAMP2) 3xHA tagged (N-terminus) I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. RAMP300000 Receptor (calcitonin) activity modifying protein 3 Insert size = 660 bp. (RAMP3) (a.k.a: receptor-activity-modifying protein 3 calcitonin receptor- like receptor activity modifying protein 3). GeneBank Accession # RAMP Proteins AY265459. RAMPs are type I transmembrane proteins with an extracellular N terminus and a cytoplasmic C terminus. RAMPs RAMP100000 Receptor (calcitonin) activity modifying protein 1 are required to transport calcitonin-receptor-like receptor (CRLR) (RAMP1) to the plasma membrane. CRLR, a receptor with seven (a.k.a: Calcitonin receptor-like receptor activity modifying protein transmembrane domains, can function as either a calcitonin- 1). GeneBank Accession # AY265457. RAMPs are required to gene-related peptide (CGRP) receptor or an adrenomedullin transport calcitonin-receptor-like receptor (CRLR) to the plasma receptor, depending on which members of the RAMP family are membrane. CRLR, a receptor with seven transmembrane expressed. In the presence of this (RAMP3) protein, CRLR domains, can function as either a calcitonin-gene-related peptide functions as an adrenomedullin receptor. Receptor (calcitonin) (CGRP) receptor or an adrenomedullin receptor, depending on activity modifying protein 3 (RAMP3) (wild type) cloned into which members of the RAMP family are expressed. In the pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI (3'). The open presence of this (RAMP1) protein, CRLR functions as a CGRP reading frame was amplified by the PCR from human Fetus receptor. The RAMP1 protein is involved in the terminal cDNA (Clontech). The insert was sequenced. Insert size= 447 glycosylation, maturation, and presentation of the CGRP RAMP30TN00 Receptor (calcitonin) activity modifying protein 3 receptor to the cell surface. Receptor (calcitonin) activity (RAMP3) 3xHA tagged (N-terminus) modifying protein 1 (RAMP1) (wild type) cloned into pcDNA3.1 (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human Lung cDNA (Clontech). The insert was sequenced. Insert size= 447. RAMP10TN00 Receptor (calcitonin) activity modifying protein 1 (RAMP1) 3xHA tagged (N-terminus)

26 cDNA Resource Center

Heterotrimeric G-proteins Alpha subunits Wild-Type Native G-protein alpha subunits pDNR-1r Native G-protein alpha subunits in pDNR-1r vector PTX-R Mutants G-protein alpha subunits resistant to Pertussis toxin due to mutation of a cysteine (C) to either isoleucine (I), glycine (G) or a serine (S), as indicated. QL Mutants G-protein alpha subunits with constitutively active phenotype due to a glutamine (Q) to leucine (L) mutation, which eliminates GTPase activity. XTP Mutants G-protein alpha subunits with dominant negative phenotype due to double glutamine (Q) to leucine (L) and aspartate (D) to asparagine (N) mutations. EE-Tagged G-protein alpha subunits epitope-tagged with an internal glutamate-glutamate epitope GT Mutants G-protein alpha subunits with dominant negative phenotype due to a glycine (G) to threonine (T) mutation QL/EE-Tagged G-protein alpha subunits with constitutively active

Alpha Subunits Wild-Type. Native G-protein alpha subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI subunits (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (669438). The insert was sequenced. GNA0L00000 G-protein alpha olf Insert size = 1085 bp. GenBank Accession # AF493893: Human G-protein alpha olf subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GNA1200000 G-protein alpha 12 (5') and Xho I (3'). The open reading frame was amplified by the GenBank Accession # AF493901: Human G-protein alpha 12 PCR from human whole brain cDNA (Clontech). The insert was subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI sequenced. Insert size = 1150 bp. (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was GNA0OA0000 G-protein alpha oA sequenced. Insert size = 1150 bp. GenBank Accession # AF493894: Human G-protein alpha oA subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GNA1300000 G-protein alpha 13 (5') and Xba I (3'). The open reading frame was amplified by the GenBank Accession # AF493902: Human G-protein alpha 13 PCR from human whole brain cDNA (Clontech). The insert was subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI sequenced. Insert size = 1070 bp. (5') and Xba I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA. The insert was sequenced. GNA0OB0000 G-protein alpha oB Insert size = 1140 bp. GenBank Accession # AF493895: Human G-protein alpha oB subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GNA1400000 G-protein alpha 14 (5') and Xba I (3'). The open reading frame was amplified by the GenBank Accession # AF493903: Human G-protein alpha 14 PCR from human whole brain cDNA (Clontech). The insert was subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI sequenced. Insert size = 1070 bp. (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (299064). The insert was sequenced. GNA0Q00000 G-protein alpha q Insert size = 1075 bp. GenBank Accession # AF493896: Human G-protein alpha q subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GNA1500000 G-protein alpha 15 (16) (5') and Xho I (3'). The open reading frame was amplified by the GenBank Accession # AF493904: Human G-protein alpha 15 PCR from IMAGE clone (46381). The insert was sequenced. subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI Insert size = 1085 bp. (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (2204272). The insert was sequenced.

Insert size = 1130 bp. GNA0SL0000 G-protein alpha S long

GenBank Accession # AF493897: Human G-protein alpha s long GNAI100000 G-protein alpha i1 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GenBank Accession # AF493905: Human G-protein alpha i1 (5') and Xho I (3'). The open reading frame was amplified by the subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI PCR from IMAGE clone (2448174). Th e insert was sequenced. (5') and Xho I (3'). The open reading frame was amplified by the Insert size = 1195 bp. PCR from IMAGE clone (1903020). The insert was sequenced.

Insert size = 1080 bp. GNA0SS0000 G-protein alpha S short

GenBank Accession # AF493898: Human G-protein alpha s GNAI200000 G-protein alpha i2 short subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at GenBank Accession # AF493906: Human G-protein alpha i2 KpnI (5') and Xho I (3'). The open reading frame was amplified subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI by the PCR from IMAGE clone (2178802). The insert was (5') and Xho I (3'). The open reading frame was amplified by the sequenced. Insert size = 1150 bp. PCR from IMAGE clone (2208983). The insert was sequenced.

Insert size = 1075 bp. GNA0Z00000 G-protein alpha z

GenBank Accession # AF493899: Human G-protein alpha z GNAI300000 G-protein alpha i3 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI GenBank Accession # AF493907: Human G-protein alpha i3 (5') and Xho I (3'). The open reading frame was amplified by the subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI PCR from human whole brain cDNA. The insert was sequenced. (5') and Xho I (3'). The open reading frame was amplified by the Insert size = 1075 bp. PCR from IMAGE clone (1687117). The insert was sequenced.

Insert size = 1075 bp. GNA1100000 G-protein alpha 11

GenBank Accession # AF493900: Human G-protein alpha 11

27 cDNA Resource Center

GNAT100000 G-protein alpha transducin rod and the 3’ RE. Internal SalI sites may be present. The insert was GenBank Accession # AF493908: Human G-protein alpha sequenced and found to be identical with GB ACC# AF493900. transducin rod subunit (wild type) cloned into pcDNA3.1+ Insert size = 1085 bp. (Invitrogen) at KpnI (5') and Xba I (3'). The open reading frame was amplified by the PCR from IMAGE clone (222533). The GNA1200001 G-protein alpha 12 (pDNR-1r) insert was sequenced. Insert size = 1060 bp. Human G-protein alpha 12 subunit (wild type) GNA1200000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAT200000 G-protein alpha transducin cone SmaI site was destroyed (*). The insert can be excised with SalI GenBank Accession # AF493909: Human G-protein alpha and the 3’ RE. Internal SalI sites may be present. The insert was transducin cone subunit (wild type) cloned into pcDNA3.1+ sequenced and found to be identical with GB ACC# AF493901. (Invitrogen) at KpnI (5') and Xho I (3'). The open reading frame Insert size = 1150 bp. was amplified by the PCR from IMAGE clone (2109792). The insert was sequenced. Insert size = 1070 bp. GNA1300001 G-protein alpha 13 (pDNR-1r) Human G-protein alpha 13 subunit (wild type) GNA1300000 cloned into pDNR-1r (Clontech) at SmaI (5') and XbaI (3'). The Alpha Subunits Wild Type in pDNR-1r. Native G- SmaI site was destroyed (*). The insert can be excised with SalI protein alpha subunits in pDNR-1r vector and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493902. GNA0OA0001 G-protein alpha oA (pDNR-1r) Insert size = 1140 bp. Human G-protein alpha oA subunit (wild type) GNA0OA0000 cloned into pDNR- 1r (Clontech) at SmaI (5') and XhoI (3'). The GNA1400001 G-protein alpha 14 (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha 14 subunit (wild type) GNA1400000 and the 3’ RE. Internal SalI sites may be present. The insert was cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The sequenced and found to be identical with GB ACC# AF493894. SmaI site was destroyed (*). The insert can be excised with SalI Insert size = 1070 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493903. GNA0OB0001 G-protein alpha oB (pDNR-1r) Insert size = 1075 bp. Human G-protein alpha oB subunit (wild type) GNA0OB0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNA1500001 G-protein alpha 15 (16) (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha 15 subunit (wild type) GNA1500000 and the 3’ RE. Internal SalI sites may be present. The insert was cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The sequenced and found to be identical with GB ACC# AF493895. SmaI site was destroyed(*). The insert can be excised with SalI Insert size = 1070 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493904. GNA0Q00001 G-protein alpha q (pDNR-1r) Insert size = 1130 bp. Human G-protein alpha q subunit (wild type) GNA0Q00000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAI100001 Human G-protein alpha i1 subunit (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha i1 subunit (wild type) GNAI100000 and the 3’ RE. Internal SalI sites may be present. The insert was cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The sequenced and found to be identical with GB ACC# AF493896. SmaI site was destroyed (*). The insert can be excised with SalI Insert size = 1085 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493905. GNA0SL0001 G-protein alpha s long (pDNR-1r) Insert size = 1080 bp. Human G-protein alpha s long subunit (wild type) GNA0SL0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAI200001 G-protein alpha i2 (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha i2 subunit (wild type) GNAI200000 and the 3’ RE. Internal SalI sites may be present. The insert was cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The sequenced and found to be identical with GB ACC# AF493897. SmaI site was destroyed (*). The insert can be excised with SalI Insert size = 1195 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493906. GNA0SS0001 G-protein alpha s short (pDNR-1r) Insert size = 1075 bp. Human G-protein alpha s short subunit (wild type) GNA0SS0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAI300001 G-protein alpha i3 (pDNR-1r) SmaI site was destroyed (* ). The insert can be excised with SalI Human G-protein alpha i3 subunit (wild type) GNAI300000 and the 3’ RE. Internal SalI sites may be present. The insert was cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The sequenced and found to be identical with GB ACC# AF493898. SmaI site was destroyed (*). The insert can be excised with SalI Insert size = 1150 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF493907. GNA0Z00001 G-protein alpha z (pDNR-1r) Insert size = 1075 bp. Human G-protein alpha z subunit (wild type) GNA0Z00000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAT100001 G-protein alpha transducin rod (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha transducin rod subunit (wild type) and the 3’ RE. Internal SalI sites may be present. The insert was GNAT100000 cloned into pDNR-1r (Clontech) at SmaI (5') and sequenced and found to be identical with GB ACC# AF493899. XbaI (3'). The SmaI site was destroyed (*). The insert can be Insert size = 1075 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GNA1100001 G-protein alpha 11 (pDNR-1r) GB ACC# AF493908. Insert size = 1060 bp. Human G-protein alpha 11 subunit (wild type) GNA1100000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNAT200001 G-protein alpha transducin cone (pDNR-1r) SmaI site was destroyed (*). The insert can be excised with SalI Human G-protein alpha transducin cone subunit (wild type)

28 cDNA Resource Center

GNAT200001 cloned into pDNR-1r (Clontech) at SmaI (5') and mutagenesis kit (Stratagene). The mutation eliminates sensitivity XhoI (3'). The SmaI site was destroyed (*). The insert can be to pertussis toxin. Insert size = 1075 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GNAI2000Q0 G-protein alpha i2 C352G GB ACC# AF493909. Insert size = 1070 bp. The C352G mutation was introduced into the human G-protein alpha i2 subunit (GNAI200000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity Alpha Subunit PTX-Resistant Mutants. G-protein to pertussis toxin. Insert size = 1075 bp. alpha subunits resistant to Pertussis toxin due to mutation of a cysteine (C) to either isoleucine (I), glycine (G) or a serine (S), as GNAI2000R0 G-protein alpha i2 C352S indicated. The C352S mutation was introduced into the human G-protein alpha i2 subunit (GNAI200000) via the Quickchange GNA0OA00P0 G-protein alpha oA C351I mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351I mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1075 bp. alpha oA subunit (GNA0OA0000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAI3000P0 G-protein alpha i3 C351I to pertussis toxin. Insert size = 1070 bp. The C351I mutation was introduced into the human G-protein alpha i3 subunit (GNAI300000) via the Quickchange GNA0OA00Q0 G-protein alpha oA C351G mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351G mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1080 bp. alpha oA subunit (GNA0OA0000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAI3000Q0 G-protein alpha i3 C351G to pertussis toxin. Insert size = 1070 bp. The C351G mutation was introduced into the human G-protein alpha i3 subunit (GNAI300000) via the Quickchange GNA0OA00R0 G-protein alpha oA C351S mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351S mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1080 bp. alpha oA subunit (GNA0OA0000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAI3000R0 G-protein alpha i3 C351S to pertussis toxin. Insert size = 1070 bp. The C351S mutation was introduced into the human G-protein alpha i3 subunit (GNAI300000) via the Quickchange GNA0OB00P0 G-protein alpha oB C351I mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351I mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1080 bp. alpha oB subunit (GNA0OB00000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT1000P0 G-protein alpha transducin rod C347I to pertussis toxin. Insert size = 1070 bp. The C347I mutation was introduced into the human G-protein alpha transducin rod subunit (GNAT100000) via the GNA0OB00Q0 G-protein alpha oB C351G Quickchange mutagenesis kit (Stratagene). The mutation The C351G mutation was introduced into the human G-protein eliminates sensitivity to pertussis toxin. Insert size = 1060 bp. alpha oB subunit (GNA0OB00000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT1000Q0 G-protein alpha transducin rod C347G to pertussis toxin. Insert size = 1070 bp. The C347G mutation was introduced into the human G-protein alpha transducin rod subunit (GNAT100000) via the GNA0OB00R0 G-protein alpha oB C351S Quickchange mutagenesis kit (Stratagene). The mutation The C351S mutation was introduced into the human G-protein eliminates sensitivity to pertussis toxin. Insert size = 1060 bp. alpha oB subunit (GNA0OB00000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT1000R0 G-protein alpha transducin rod C347S to pertussis toxin. Insert size = 1070 bp. The C347S mutation was introduced into the human G-protein alpha transducin rod subunit (GNAT100000) via the GNAI1000P0 G-protein alpha i1 C351I Quickchange mutagenesis kit (Stratagene). The mutation The C351I mutation was introduced into the human G-protein eliminates sensitivity to pertussis toxin. Insert size = 1060 bp. alpha i1 subunit (GNAI100000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT2000P0 G-protein alpha transducin cone C351I to pertussis toxin. Insert size = 1080 bp. The C351I mutation was introduced into the human G-protein alpha transducin cone (GNAT200000) via the Quickchange GNAI1000Q0 G-protein alpha i1 C351G mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351G mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1070 bp. alpha i1 subunit (GNAI100000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT2000Q0 G-protein alpha transducin cone C351G to pertussis toxin. Insert size = 1080 bp. The C351G mutation was introduced into the human G-protein alpha transducin cone (GNAT200000) via the Quickchange GNAI1000R0 G-protein alpha i1 C351S mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C351S mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1070 bp. alpha i1 subunit (GNAI100000) via the Quickchange mutagenesis kit (Stratagene). The mutation eliminates sensitivity GNAT2000R0 G-protein alpha transducin cone C351S to pertussis toxin. Insert size = 1080 bp. The C351S mutation was introduced into the human G-protein alpha transducin cone (GNAT200000) via the Quickchange GNAI2000P0 G-protein alpha i2 C352I mutagenesis kit (Stratagene). The mutation eliminates sensitivity The C352I mutation was introduced into the human G-protein to pertussis toxin. Insert size = 1070 bp. alpha i2 subunit (GNAI200000) via the Quickchange

29 cDNA Resource Center

Alpha Subunits QL Mutants. G-protein alpha subunits GNA14000C0 G-protein alpha 14 Q205L with constitutively active phenotype due to a glutamine (Q) to The Q205L mutation was introduced into the human G-protein leucine (L) mutation, which eliminates GTPase activity. alpha 14 (GNA1400000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in GNA0L000C0 G-protein alpha olf Q214L a constitutively active phenotype. Insert size = 1075 bp. The Q214L mutation was introduced into the human G-protein alpha olf (GNA0L00000) via the Quickchange mutagenesis kit GNA15000C0 G-protein alpha 15 Q212L (Stratagene). The mutation reduces GTPase activity resulting in The Q212L mutation was introduced into the human G-protein a constitutively active phenotype. Insert size = 1150 bp. alpha 15 (GNA1500000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in GNA0OA00C0 G-protein alpha oA Q205L a constitutively active phenotype. Insert size = 1130 bp. The Q205L mutation was introduced into the human G-protein alpha oA (GNA0OA0000) via the Quickchange mutagenesis kit GNAI1000C0 G-protein alpha i1 Q204L (Stratagene). The mutation reduces GTPase activity resulting in The Q204L mutation was introduced into the human G-protein a constitutively active phenotype. Insert size = 1070 bp. alpha i1 subunit (GNAI100000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase GNA0OB00C0 G-protein alpha oB Q205L activity resulting in a constitutively active phenotype. Insert size The Q205L mutation was introduced into the human G-protein = 1080 bp. alpha oB (GNA0OB0000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in GNAI2000C0 G-protein alpha i2 Q205L a constitutively active phenotype. Insert size = 1070 bp. The Q205L mutation was introduced into the human G-protein alpha i2 subunit (GNAI200000) via the Quickchange GNA0Q000C0 G-protein alpha q Q209L mutagenesis kit (Stratagene). The mutation reduces GTPase The Q209L mutation was introduced into the human G-protein activity resulting in a constitutively active phenotype. Insert size alpha q subunit (GNA0Q00000) via the Quickchange = 1075 bp. mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active phenotype. Insert size GNAI3000C0 G-protein alpha i3 Q204L = 1085 bp. The Q204L mutation was introduced into the human G-protein alpha i3 subunit (GNAI300000) via the Quickchange GNA0SL00C0 G-protein alpha s long Q227L mutagenesis kit (Stratagene). The mutation reduces GTPase The Q227L mutation was introduced into the human G-protein activity resulting in a constitutively active phenotype. Insert size alpha s long subunit (GNA0SL00000) via the Quickchange = 1075 bp. mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active phenotype. Insert size GNAT1000C0 G-protein alpha transducin rod Q200L = 1195 bp. The Q200L mutation was introduced into the human G-protein alpha transducin rod (GNAT100000) via the Quickchange GNA0SS00C0 G-protein alpha s short Q213L mutagenesis kit (Stratagene). The mutation reduces GTPase The Q213L mutation was introduced into the human G-protein activity resulting in a constitutively active phenotype. Insert size alpha s short subunit (GNA0SS0000) via the Quickchange = 1060 bp. mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active phenotype. Insert size GNAT2000C0 G-protein alpha transducin cone Q204L = 1070 bp. The Q204L mutation was introduced into the human G-protein alpha transducin cone subunit (GNAT200000) via the GNA0Z000C0 G-protein alpha z Q205L Quickchange mutagenesis kit (Stratagene). The mutation The Q205L mutation was introduced into the human G-protein reduces GTPase activity resulting in a constitutively active alpha z (GNA0Z00000) via the Quickchange mutagenesis kit phenotype. Insert size = 1070 bp. (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active phenotype. Insert size = 1075 bp. Alpha Subunit XTP Mutants. G-protein alpha subunits GNA11000C0 G-protein alpha 11 Q209L with dominant negative phenotype due to double glutamine (Q) The Q209L mutation was introduced into the human G-protein to leucine (L) and aspartate (D) to asparagine (N) mutations. alpha 11 subunit (GNA1100000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase GNA0L000X0 G-protein alpha olf Q214L/D282N activity resulting in a constitutively active phenotype. Insert size The Q214L and D282N mutations were introduced into the = 1085 bp. human G-protein alpha olf subunit (GNA0L00000) via the Quickchange mutagenesis kit (Stratagene). The mutations GNA12000C0 G-protein alpha 12 Q231L confer a preference for xanthine nucleotide binding. Insert size = The Q231L mutation was introduced into the human G-protein 1150 bp. alpha 12 (GNA1200000) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in GNA0OA00X0 G-protein alpha oA Q205L/D273N a constitutively active phenotype. Insert size = 1150 bp. The Q205L and D273N mutations were introduced into the human G-protein alpha oA subunit (GNA0OA0000) via the GNA13000C0 G-protein alpha 13 Q226L Quickchange mutagenesis kit (Stratagene). The mutations The Q226L mutation was introduced into the human G-protein confer a preference for xanthine nucleotide binding. Insert size = alpha 13 subunit (GNA1300000) via the Quickchange 1070 bp. mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active phenotype. Insert size GNA0OB00X0 G-protein alpha oB Q205L/D273N = 1140 bp. The Q205L and D273N mutations were introduced into the human G-protein alpha oB subunit (GNA0OB0000) via the Quickchange mutagenesis kit (Stratagene). The mutations

30 cDNA Resource Center

confer a preference for xanthine nucleotide binding. Insert size = 1070 bp. GNAI2000X0 G-protein alpha i2 Q205L/D273N The Q205L and D273N mutations were introduced into the GNA0Q000X0 G-protein alpha q Q209L/D277N human G-protein alpha i2 subunit (GNAI200000) via the The Q209L and D277N mutations were introduced into the Quickchange mutagenesis kit (Stratagene). The mutations human G-protein alpha q subunit (GNA0Q00000) via the confer a preference for xanthine nucleotide binding. Insert size = Quickchange mutagenesis kit (Stratagene). The mutations 1075 bp. confer a preference for xanthine nucleotide binding. Insert size = 1085 bp. GNAI3000X0 G-protein alpha i3 Q204L/D272N The Q204L and D272N mutations were introduced into the GNA0SL00X0 G-protein alpha S long Q227L/D295N human G-protein alpha i3 subunit (GNAI300000) via the The Q213L and D281N mutations were introduced into the Quickchange mutagenesis kit (Stratagene). The mutations human G-protein alpha s short subunit (GNA0SS0000) via the confer a preference for xanthine nucleotide binding. Insert size = Quickchange mutagenesis kit (Stratagene). The mutations 1075 bp. confer a preference for xanthine nucleotide binding. Insert size = 1150 bp. GNAT1000X0 G-protein alpha transducin rod Q200L/D268N The Q200L and D268N mutations were introduced into the GNA0SS00X0 G-protein alpha S short Q213L/D281N human G-protein alpha transducin rod subunit (GNAT100000) The Q213L and D281N mutations were introduced into the via the Quickchange mutagenesis kit (Stratagene). The human G-protein alpha s short subunit (GNA0SS0000) via the mutations confer a preference for xanthine nucleotide binding. Quickchange mutagenesis kit (Stratagene). The mutations Insert size = 1060 bp. confer a preference for xanthine nucleotide binding. Insert size = 1150 bp. GNAT2000X0 G-protein alpha transducin cone Q204L/D272N The Q204L and D272N mutations were introduced into the GNA0Z000X0 G-protein alpha z Q205L/D273N human G-protein alpha transducin cone subunit (GNAT200000) The Q205L and D273N mutations were introduced into the via the Quickchange mutagenesis kit (Stratagene). The human G-protein alpha z subunit (GNA0Z00000) via the mutations confer a preference for xanthine nucleotide binding. Quickchange mutagenesis kit (Stratagene). The mutations Insert size = 1070 bp. confer a preference for xanthine nucleotide binding. Insert size = 1075 bp. Alpha Subunits EE-Tagged. G-protein alpha subunits GNA11000X0 G-protein alpha 11 Q209L/D277N epitope-tagged with an internal glutamate-glutamate epitope. The Q209L and D277N mutations were introduced into the human G-protein alpha 11 subunit (GNA1100000) via the GNA0L0EI00 G-protein alpha olf EE tagged (internal) Quickchange mutagenesis kit (Stratagene). The mutations An internal Glu-Glu epitope tag (see reference above) was confer a preference for xanthine nucleotide binding. Insert size = introduced into the human G-protein alpha olf subunit (wild 1085 bp. type;GNA0L00000) via the Quickchange mutagenesis kit (Stratagene). Residues DYTPTD (176-181) were mutated to GNA12000X0 G-protein alpha 12 Q231L/D299N EYMPTE. The Q231L and D299N mutations were introduced into the human G-protein alpha 12 (GNA1200000) via the Quickchange GNA0OAEI00 G-protein alpha oA EE tagged (internal) mutagenesis kit (Stratagene). The mutations confer a preference An internal Glu-Glu epitope tag (see reference above) was for xanthine nucleotide binding. Insert size = 1150 bp. introduced into the human G-protein alpha oA subunit (wild type;GNA0OA000) via the Quickchange mutagenesis kit GNA13000X0 G-protein alpha 13 Q226L/D294N (Stratagene). Residues DYQPTE (167-172) were mutated to The Q226L and D294N mutations were introduced into the EYMPTE. human G-protein alpha 13 subunit (GNA1300000) via the Quickchange mutagenesis kit (Stratagene). The mutations GNA0OBEI00 G-protein alpha oB EE-tagged (internal) confer a preference for xanthine nucleotide binding. Insert size = An internal Glu-Glu epitope tag was introduced into the human 1140 bp. G-protein alpha oB subunit (wild type;GNA0OB0000) via the Quickchange mutagenesis kit (Stratagene). Residues DYQPTE GNA14000X0 G-protein alpha 14 Q205L/D273N (167-172) were mutated to EYMPTE. The Q205L and D273N mutations were introduced into the human G-protein alpha 14 (GNA1400000) via the Quickchange GNA0Q0EI00 G-protein alpha q EE tagged (internal) mutagenesis kit (Stratagene). The mutations confer a preference An internal Glu-Glu epitope tag (see reference above) was for xanthine nucleotide binding. Insert size = 1075 bp. introduced into the human G-protein alpha q subunit (wild type;GNA0Q0000) via the Quickchange mutagenesis kit GNA15000X0 G-protein alpha 15(16) Q212L/D280N (Stratagene). Residues AYLPTQ (171-176) were mutated to The Q212L and D280N mutations were introduced into the EYMPTE. human G-protein alpha 15 subunit (GNA1500000) via the Quickchange mutagenesis kit (Stratagene). The mutations GNA0SLEI00 G-protein alpha S long EE tagged (internal) confer a preference for xanthine nucleotide binding. Insert size = An internal Glu-Glu epitope tag (see reference above) was 1130 bp. introduced into the human G-protein alpha s long subunit (wild type;GNA0SL0000) via the Quickchange mutagenesis kit GNAI1000X0 G-protein alpha i1 Q204L/D272N (Stratagene). Residues DYVPSD (189-194) were mutated to The Q204L and D272N mutations were introduced into the EYMPTE. human G-protein alpha i1 subunit (GNAI100000) via the Quickchange mutagenesis kit (Stratagene). The mutations GNA0SSEI00 G-protein alpha S short EE tagged (internal) confer a preference for xanthine nucleotide binding. Insert size = An internal Glu-Glu epitope tag (see reference above) was 1080 bp. introduced into the human G-protein alpha olf subunit (wild

31 cDNA Resource Center

type;GNA0SS0000) via the Quickchange mutagenesis kit subunit (wild type;GNAT100000) via the Quickchange (Stratagene). Residues DYVPSD (175-180) were mutated to mutagenesis kit (Stratagene). Residues GYVPTE (162-167) EYMPTE. were mutated to EYMPTE.

GNA0Z0EI00 G-protein alpha z EE tagged (internal) GNAT20EI00 G-protein alpha transducin cone EE tagged An internal Glu-Glu epitope tag (see reference above) was (internal) introduced into the human G-protein alpha transducin rod An internal Glu-Glu epitope tag (see reference above) was subunit (wild type;GNA0Z00000) via the Quickchange introduced into the human G-protein alpha transducin cone mutagenesis kit (Stratagene). Residues DYIPTV (167-172) were subunit (wild type;GNAT200000) via the Quickchange mutated to EYMPTE. mutagenesis kit (Stratagene). Residues EYLPSE (166-171) were mutated to EYMPTE. GNA110EI00 G-protein alpha 11 EE tagged (internal) An internal Glu-Glu epitope tag (see reference above) was introduced into the human G-protein alpha 11 subunit (wild Alpha Subunits GT Mutants. G-protein alpha subunits type;GNA110000) via the Quickchange mutagenesis kit with dominant negative phenotype due to a glycine (G) to (Stratagene). Residues GYLPTQ (171-176) were mutated to threonine (T) mutation EYMPTE. GNA0OA00T0 G-protein alpha oA G203T GNA120EI00 G-protein alpha 12 EE tagged (internal) The G203T mutation was introduced into the human G-protein An internal Glu-Glu epitope tag was introduced into the human alpha oA subunit (GNA0OA0000) via the Quickchange G-protein alpha 12 subunit (wild type;GNA1200000) via the mutagenesis kit (Stratagene). The mutation produces a Quickchange mutagenesis kit (Stratagene). Residues NYFPSK putatively dominant-negative phenotype. Insert size = 1070 bp. (193-198) were mutated to EYMPTE.

GNA130EI00 G-protein alpha 13 EE tagged (internal) GNA0OB00T0 G-protein alpha oB G203T An internal Glu-Glu epitope tag (see reference above) was The G203T mutation was introduced into the human G-protein introduced into the human G-protein alpha 13 subunit (wild alpha oB subunit (GNA0OB0000) via the Quickchange type;GNA130000) via the Quickchange mutagenesis kit mutagenesis kit (Stratagene). The mutation produces a (Stratagene). Residues DYIPSQ (188-193) were mutated to putatively dominant-negative phenotype. Insert size = 1070 bp. EYMPTE. GNAI1000T0 G-protein alpha i1 G202T GNA140EI00 G-protein alpha 14 EE tagged (internal) The G202T mutation was introduced into the human G-protein An internal Glu-Glu epitope tag (see reference above) was alpha i1 subunit (GNAI100000) via the Quickchange introduced into the human G-protein alpha 14 subunit (wild mutagenesis kit (Stratagene). The mutation produces a type;GNA1400000) via the Quickchange mutagenesis kit putatively dominant-negative phenotype. Insert size = 1075 bp. (Stratagene). Residues sfvptq (167-172) were mutated to EYMPTE. GNAI2000T0 G-protein alpha i2 G203T The G203T mutation was introduced into the human G-protein GNA150EI00 G-protein alpha 15 EE tagged (internal) alpha i2 subunit (GNAI200000) via the Quickchange An internal Glu-Glu epitope tag (see reference above) was mutagenesis kit (Stratagene). The mutation produces a introduced into the human G-protein alpha 15 subunit (wild putatively dominant-negative phenotype. Insert size = 1075 bp. type;GNA1500000) via the Quickchange mutagenesis kit (Stratagene). Residues GYVPTA (174-179) were mutated to GNAI3000T0 G-protein alpha i3 G202T EYMPTE. The G202T mutation was introduced into the human G-protein alpha i3 subunit (GNAI300000) via the Quickchange GNAI10EI00 G-protein alpha i1 EE tagged (internal) mutagenesis kit (Stratagene). The mutation produces a An internal Glu-Glu epitope tag (see reference above) was putatively dominant-negative phenotype. Insert size = 1075 bp. introduced into the human G-protein alpha olf subunit (wild type;GNAI100000) via the Quickchange mutagenesis kit (Stratagene). Residues NYIPTQ (166-171) were mutated to Alpha Subunits QL/EE-Tagged. G-protein alpha EYMPTE. subunits with constitutively active phenotype and an internal

glutamate-glutamate epitope GNAI20EI00 G-protein alpha i2 EE tagged (internal)

An internal Glu-Glu epitope tag (see reference above) was GNA0L0EIC0 G-protein alpha olf Q214L EE-tagged (internal) introduced into the human G-protein alpha i2 subunit (wild The Q214L mutation was introduced into the internally Glu-Glu type;GNAI200000) via the Quickchange mutagenesis kit epitope tagged human G-protein alpha olf (GNA0L0EI00) via the (Stratagene). Residues DYIPTQ (167-172) were mutated to Quickchange mutagenesis kit (Stratagene). The mutation EYMPTE. reduces GTPase activity resulting in a constitutively active

phenotype. Insert size = 1150 bp. GNAI30EI00 G-protein alpha i3 EE tagged (internal)

An internal Glu-Glu epitope tag (see reference above) was GNA0OAEIC0 G-protein alpha oA Q205L EE-tagged introduced into the human G-protein alpha i3 subunit (wild (internal) type;GNAI300000) via the Quickchange mutagenesis kit The Q205L mutation was introduced into the internally Glu-Glu (Stratagene). Residues nYIPTQ (166-171) were mutated to epitope tagged human G-protein alpha oA (GNA0OAEI00) via EYMPTE. the Quickchange mutagenesis kit (Stratagene). The mutation

reduces GTPase activity resulting in a constitutively active GNAT10EI00 G-protein alpha transducin rod EE tagged phenotype. Insert size = 1070 bp. (internal)

An internal Glu-Glu epitope tag (see reference above) was introduced into the human G-protein alpha transducin rod

32 cDNA Resource Center

GNA0OBEIC0 G-protein alpha oB Q205L EE-tagged Quickchange mutagenesis kit (Stratagene). The mutation (internal) reduces GTPase activity resulting in a constitutively active The Q205L mutation was introduced into the internally Glu-Glu phenotype. Insert size = 1140 bp. epitope tagged human G-protein alpha oB (GNA0OBEI00) via the Quickchange mutagenesis kit (Stratagene). The mutation GNA140EIC0 G-protein alpha 14 Q205L EE-tagged (internal) reduces GTPase activity resulting in a constitutively active The Q205L mutation was introduced into the internally Glu-Glu phenotype. Insert size = 1070 bp. epitope tagged human G-protein alpha 14 (GNA140EI00) via the Quickchange mutagenesis kit (Stratagene). The mutation GNA0Q0EIC0 G-protein alpha q Q209L EE-tagged (internal) reduces GTPase activity resulting in a constitutively active The Q209L mutation was introduced into the internally Glu-Glu phenotype. Insert size = 1075 bp. epitope tagged human G-protein alpha q (GNA0Q0EI00) via the Quickchange mutagenesis kit (Stratagene). The mutation GNA150EIC0 G-protein alpha 15 Q212L EE-tagged (internal) reduces GTPase activity resulting in a constitutively active The Q212L mutation was introduced into the internally Glu-Glu phenotype. Insert size = 1085 bp. epitope tagged human G-protein alpha 15 (GNA150EI00) via the Quickchange mutagenesis kit (Stratagene). The mutation GNA0SLEIC0 G-protein alpha S long Q227L EE-tagged reduces GTPase activity resulting in a constitutively active (internal) phenotype. Insert size = 1130 bp. The Q227L mutation was introduced into the internally Glu-Glu epitope tagged human G-protein alpha S long (GNA0SLEI00) via GNAI10EIC0 G-protein alpha i1 Q204L EE-tagged (internal) the Quickchange mutagenesis kit (Stratagene). The mutation The Q204L mutation was introduced into the internally Glu-Glu reduces GTPase activity resulting in a constitutively active epitope tagged human G-protein alpha i1 (GNAI10EI00) via the phenotype. Insert size = 1195 bp. Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in a constitutively active GNA0SSEIC0 G-protein alpha S short Q213L EE-tagged phenotype. Insert size = 1080 bp. (internal) The Q213L mutation was introduced into the internally Glu-Glu GNAI20EIC0 G-protein alpha i2 Q205L EE-tagged (internal) epitope tagged human G-protein alpha S short (GNA0SSEI00) The Q205L mutation was introduced into the internally Glu-Glu via the Quickchange mutagenesis kit (Stratagene). The mutation epitope tagged human G-protein alpha i2 (GNAI20EI00) via the reduces GTPase activity resulting in a constitutively active Quickchange mutagenesis kit (Stratagene). The mutation phenotype. Insert size = 1150 bp. reduces GTPase activity resulting in a constitutively active phenotype. Insert size = 1075 bp. GNA0Z0EIC0 G-protein alpha z Q205L EE-tagged (internal) The Q205L mutation was introduced into the internally Glu-Glu GNAI30EIC0 G-protein alpha i3 Q204L EE-tagged (internal) epitope tagged human G-protein alpha z (GNA0Z0EI00) via the The Q204L mutation was introduced into the internally Glu-Glu Quickchange mutagenesis kit (Stratagene). The mutation epitope tagged human G-protein alpha i3 (GNAI30EI00) via the reduces GTPase activity resulting in a constitutively active Quickchange mutagenesis kit (Stratagene). The mutation phenotype. Insert size = 1075 bp. reduces GTPase activity resulting in a constitutively active phenotype. Insert size = 1075 bp. GNA110EIC0 G-protein alpha 11 Q209L EE-tagged (internal) The Q209L mutation was introduced into the internally Glu-Glu GNAT10EIC0 G-protein alpha transducin rod Q200L EE- epitope tagged human G-protein alpha 11 (GNA110EI00) via the tagged (internal) Quickchange mutagenesis kit (Stratagene). The mutation The Q200L mutation was introduced into the internally Glu-Glu reduces GTPase activity resulting in a constitutively active epitope tagged human G-protein alpha transducin rod phenotype. Insert size = 1085 bp. (GNAT10EI00) via the Quickchange mutagenesis kit (Stratagene). The mutation reduces GTPase activity resulting in GNA120EIC0 G-protein alpha 12 Q231L EE-tagged (internal) a constitutively active phenotype. Insert size = 1060 bp. The Q231L mutation was introduced into the internally Glu-Glu epitope tagged human G-protein alpha 12 (GNA120EI00) via the GNAT20EIC0 G-protein alpha transducin cone Q204L EE- Quickchange mutagenesis kit (Stratagene). The mutation tagged (internal) reduces GTPase activity resulting in a constitutively active The Q204L mutation was introduced into the internally Glu-Glu phenotype. Insert size = 1150 bp. epitope tagged human G-protein alpha transducin cone (GNAT20EI00) via the Quickchange mutagenesis kit GNA130EIC0 G-protein alpha 13 Q226L EE-tagged (internal) (Stratagene). The mutation reduces GTPase activity resulting in The Q226L mutation was introduced into the internally Glu-Glu a constitutively active phenotype. Insert size = 1070 bp. epitope tagged human G-protein alpha 13 (GNA130EI00) via the

33 cDNA Resource Center

Heterotrimeric G-proteins Beta subunits Wild-Type Native G-protein beta subunits FLAG-Tagged Native G-protein beta subunits with an N-terminal FLAG epitope tag pDNR-1r Native G-protein beta subunits in pDNR-1r vector

Beta Wild-Type. Native G-protein beta subunits Beta FLAG-Tagged. Native G-protein beta subunits with GNB0100000 G-protein beta 1 an N-terminal FLAG epitope tag GenBank Accession # AF501882: Human G-protein beta 1 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I GNB010FN00 G-protein beta 1 FLAG-tagged (N-terminus) (5') and Xho I (3'). The open reading frame was amplified by the N-terminal FLAG-tagged human G-protein beta 1 subunit (wild PCR from IMAGE clone (2001658). The insert was sequenced. type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I Insert size = 1025 bp. (3'). The open reading frame was derived from GNB0100000 by the PCR. Insert size = 1050 bp. GNB0200000 G-protein beta 2 GenBank Accession # AF501883: Human G-protein beta 2 GNB020FN00 G-protein beta 2 FLAG-tagged (N-terminus) subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I N-terminal FLAG-tagged human G-protein beta 2 subunit (wild (5') and Xho I (3'). The open reading frame was amplified by the type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I PCR from IMAGE clone (1962114). The insert was sequenced. (3'). The open reading frame was derived from GNB0200000 by Insert size = 1025 bp. the PCR. Insert size = 1050 bp.

GNB0300000 G-protein beta 3 GNB030FN00 G-protein beta 3 FLAG-tagged (N-terminus) GenBank Accession # AF501884: Human G-protein beta 3 N-terminal FLAG-tagged human G-protein beta 3 subunit (wild subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I (5') and Xho I (3'). The open reading frame was amplified by the (3'). The open reading frame was derived from GNB0300000 by PCR from human whole brain cDNA (Clontech). The insert was the PCR. Insert size = 1050 bp. sequenced. Insert size = 1025 bp. GNB03SFN00 G-protein beta 3S FLAG-tagged (N-terminus) GNB03S0000 G-protein beta 3S N-terminal FLAG-tagged human G-protein beta 3s subunit (wild Human G-protein beta 3s subunit (splice variant) cloned into type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I pcDNA3.1+ (Invitrogen) at EcoR I (5') and Not I (3'). The open (3'). The open reading frame was derived from GNB03S0000 by reading frame was constructed by the PCR from G-protein beta the PCR. Insert size = 930 bp. 3 wt. The insert was sequenced. Insert size = 900 bp. GNB040FN00 G-protein beta 4 FLAG-tagged (N-terminus) GNB0400000 G-protein beta 4 N-terminal FLAG-tagged human G-protein beta 4 subunit (wild GenBank Accession # AF300648: Human G-protein beta 4 type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (3'). The open reading frame was derived from GNB0400000 by (5') and Xho I (3'). The open reading frame was amplified by the the PCR. Insert size = 1050 bp. PCR from human whole brain cDNA (Clontech). The insert was sequenced and the translated protein found to be similar to GNB050FN00 G-protein beta 5 FLAG-tagged (N-terminus) mouse beta 4. Insert size = 1025 bp. N-terminal FLAG-tagged human G-protein beta 5 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I GNB040000M G-protein beta 4 (mouse) (3'). The open reading frame was derived from GNB0500000 by GenBank Accession # AF501886: Mouse G-protein beta 4 the PCR. Insert size = 1100 bp. subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was amplified by the GNB05LFN00 G-protein beta 5L FLAG-tagged (N-terminus) PCR from IMAGE clone (457406). The insert was sequenced. N-terminal FLAG-tagged human G-protein beta 5L subunit (wild Insert size = 1040 bp. type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and Xho I (3'). The open reading frame was derived from GNB05L0000 by GNB0500000 G-protein beta 5 the PCR. Insert size = 1225 bp. GenBank Accession # AF501885: Human G-protein beta 5 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was amplified by the Beta Wild-Type in pDNR-1r. Native G-protein beta PCR from IMAGE clone (37537). The insert was sequenced. subunits in pDNR-1r vector Insert size = 1070 bp. GNB0100001 G-protein beta 1 (pDNR-1r) GNB05L0000 G-protein beta 5L Human G-protein beta 1 subunit (wild type) GNB0100000 GenBank Accession # AF300650: Human G-protein beta 5L cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I SmaI site was destroyed(*). The insert can be excised with SalI (5') and Xho I (3'). The open reading frame was amplified by the and the 3’ RE. Internal SalI sites may be present. The insert was PCR from human retinal cDNA (Clontech). N- terminal primer sequenced and found to be identical with GB ACC# AF501882. sequence derived from EST clone AA317427. The insert was Insert size = 1025 bp. sequenced. 41 out of the 42 additional predicted N-terminal amino acids are identical to mouse G-beta 5L (M87286). Insert GNB0200001 G-protein beta 2 (pDNR-1r) size = 1200 bp. Human G-protein beta 2 subunit (wild type) GNB0200000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The

34 cDNA Resource Center

SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC#AF501883. Insert size = 1025 bp.

GNB0300001 G-protein beta 3 (pDNR-1r) Human G-protein beta 3 subunit (wild type) GNB0300000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC# AF501884. Insert size = 1025 bp.

GNB03S0001 G-protein beta 3S (pDNR-1r) Human G-protein beta 3s subunit (splice variant) GNB03S0000 cloned into pDNR-1r (Clontech) at SmaI (5') and HindIII (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical to human beta 3 wt with a 41 amino acid deletion as described by Siffert et al. Nature Genetics 18: 45-48 (1998). Insert size = 900 bp.

GNB0400001 G-protein beta 4 (pDNR-1r) Human G-protein beta 4 subunit (wild type) GNB0400000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and the translated protein found to be similar to mouse beta 4. Insert size = 1025 bp.

GNB0400002 mouse G-protein beta 4 (pDNR-1r) Mouse G-protein beta 4 subunit (wild type) GNB040000M cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and compared to GB ACC# AF300648. Insert size = 1040 bp.

GNB05L0001 G-protein beta 5L (pDNR-1r) Human G-protein beta 5L subunit (wild type) GNB05L0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and bases 127- 1188 were identical with GB ACC# AF300650. 41 out of the 42 additional predicted N-terminal amino acids are identical to mouse Gbeta 5L (M87286). Insert size = 1200 bp. The insert was sequenced and found to be identical with GB ACC# M31328 with the following exceptions: A339->G, T657->A and C825->T (all mutations are silent). Insert size = 1025 bp.

35 cDNA Resource Center

Heterotrimeric G-proteins Gamma subunits

Wild-Type Native G-protein gamma subunits HA-Tagged Native G-protein gamma subunits with an N-terminal hemagglutinin epitope tag pDNR-1r Native G-protein gamma subunits in pDNR-1r vector

Gamma Wild-Type. Native G-protein gamma subunits GNG1000000 G-protein gamma 10 GenBank Accession # AF493877: Human G-protein gamma 10 GNG0100000 G-protein gamma 1 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Hind III GenBank Accession # AF493869: Human G-protein gamma (5') and Xho I (3'). The open reading frame was amplified by the subunit 1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I PCR from IMAGE clone (290588). The insert was sequenced. (5') and Xho I (3'). The open reading frame was amplified by the Insert size = 220 bp. PCR from IMAGE clone (381155). The insert was sequenced. Insert size = 225 bp. GNG1100000 G-protein gamma 11 GenBank Accession # AF493878: Human G-protein gamma 11 GNG0200000 G-protein gamma 2 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Hind III GenBank Accession # AF493870: Human G-protein gamma 2 (5') and Xho I (3'). The open reading frame was amplified by the subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I PCR from IMAGE clone (550119). The insert was sequenced. (5') and Xho I (3'). The open reading frame was amplified by the Insert size = 235 bp. PCR from IMAGE clone (1408502). Predicted amino acid sequence is identical to bovine G-protein gamma 2 (M37183). GNG1200000 G-protein gamma 12 The insert was sequenced. Insert size = 220 bp. GenBank Accession # AF493879: Human G-protein gamma 12 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I GNG0300000 G-protein gamma 3 (5') and Xho I (3'). The open reading frame was amplified by the GenBank Accession # AF493871: Human G-protein gamma 3 PCR from IMAGE clone (306388). The insert was sequenced. subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I Insert size = 227 bp. (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (632992). The insert was sequenced. GNG1300000 G-protein gamma 13 Insert size = 232 bp. GenBank Accession # AF493880: Human G-protein gamma 13 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I GNG0400000 G-protein gamma 4 (5') and Xho I (3'). The open reading frame was amplified by the GenBank Accession # AF493872: Human G-protein gamma 4 PCR from IMAGE clone (178213). The predicted amino acid subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I sequence is identical to that reported by Huang et al. (1999). (5') and Xho I (3'). The open reading frame was amplified by the Insert size = 213 bp. PCR from IMAGE clone (2340744). The insert was sequenced. Insert size = 232 bp. Gamma HA-Tagged. Native G-protein gamma subunits GNG0500000 G-protein gamma 5 with an N-terminal hemagglutinin epitope tag GenBank Accession # AF493873: Human G-protein gamma 5 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I GNG010HN00 G-protein gamma 1 HA-tagged (N-terminus) (5') and Xho I (3'). The open reading frame was amplified by the N-terminal hemagglutinin tagged human G-protein gamma 1 PCR from IMAGE clone (2453124). The insert was sequenced. subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI Insert size = 210 bp. (5') and Xho I (3'). The open reading frame was derived from GNG0100000 by the PCR. Insert size = 250 bp. GNG0700000 G-protein gamma 7 GenBank Accession # AF493874: Human G-protein gamma 7 GNG020HN00 G-protein gamma 2 HA-tagged (N-terminus) subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I N-terminal hemagglutinin tagged human G-protein gamma 2 (5') and Xho I (3'). The open reading frame was amplified by the subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI PCR from IMAGE clone (190059). The insert was sequenced. (5') and Xho I (3'). The open reading frame was derived from Insert size = 210 bp. GNG0200000 by the PCR. Insert size = 250 bp.

GNG0800000 G-protein gamma 8 GNG030HN00 G-protein gamma 3 HA-tagged (N-terminus) GenBank Accession # AF493875: Human G-protein gamma 8 N-terminal hemagglutinin tagged human G-protein gamma 3 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame was amplified by the (5') and Xho I (3'). The open reading frame was derived from PCR from IMAGE clone (2207401). The insert was sequenced. GNG0300000 by the PCR. Insert size = 255 bp. Insert size = 223 bp. GNG040HN00 G-protein gamma 4 HA tagged (N-terminus) GNG0900000 G-protein gamma 9 N-terminal hemagglutinin tagged human G-protein gamma 4 GenBank Accession # AF493876: Human G-protein gamma 9 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame was derived from (5') and Xho I (3'). The open reading frame was amplified by the GNG0400000 by the PCR. Insert size = 250 bp. PCR from IMAGE clone (1535311). The insert was sequenced. Insert size = 219 bp. Predicted amino acid sequence is 86% GNG050HN00 G-protein gamma 5 HA tagged (N-terminus) identical to bovine G-protein gamma 9 (U20085). N-terminal hemagglutinin tagged human G-protein gamma 5 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoR I

36 cDNA Resource Center

(5') and Xho I (3'). The open reading frame was derived from sequenced and found to be identical with GB ACC#AF493871. GNG0500000 by the PCR. Insert size = 250 bp. Insert size = 232 bp.

GNG070HN00 G-protein gamma 7 HA-tagged (N-terminus) GNG0400001 G-protein gamma 4 (pDNR-1r) N-terminal hemagglutinin tagged human G-protein gamma 7 Human G-protein gamma 4 subunit (wild type) GNG0100000 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI cloned into pDNR- 1R (Clontech) at SmaI (5') and XhoI (3'). The (5') and Xho I (3'). The open reading frame was derived from SmaI site was destroyed(*). The insert can be excised with SalI GNG0700000 by the PCR. Insert size = 240 bp. and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC#AF493872. GNG080HN00 G-protein gamma 8 HA tagged (N-terminus) Insert size = 232 bp. N-terminal hemagglutinin tagged human G-protein gamma 8 subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI GNG0500001 G-protein gamma 5 (pDNR-1r) (5') and Xho I (3'). The open reading frame was derived from Human G-protein gamma 5 subunit (wild type) GNG0100000 GNG0800000 by the PCR. Insert size = 260 bp. cloned into pDNR-1r (Clontech) at SmaI(5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI GNG090HN00 G-protein gamma 9 HA tagged (N-terminus) and the 3’ RE. The insert was sequenced and found to be N-terminal hemagglutinin tagged human G-protein gamma 9 identical with GB ACC# AF493873. Insert size = 210 bp. subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was derived from GNG0700001 G-protein gamma 7 (pDNR-1r) GNG0900000 by the PCR. Insert size = 250 bp. Human G-protein gamma 7 subunit (wild type) GNG0700000 cloned into pDNR- 1r (Clontech) at SmaI (5') and XhoI (3'). The GNG100HN00 G-protein gamma 10 HA tagged (N-terminus) SmaI site was destroyed(*). The insert can be excised with SalI N-terminal hemagglutinin tagged human G-protein gamma 10 and the 3’RE. Internal SalI sites may be present. The insert was subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI sequenced and found to be identical with GB ACC#AF493874. (5') and Xho I (3'). The open reading frame was derived from Insert size = 210 bp. GNG1000000 by the PCR. Insert size = 250 bp. GNG0800001 G-protein gamma 8 (pDNR-1r) GNG110HN00 G-protein gamma 11 HA tagged (N-terminus) Human G-protein gamma 8 subunit (wild type) GNG0800001 N-terminal hemagglutinin tagged human G-protein gamma 11 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI SmaI site was destroyed(*). The insert can be excised with SalI (5') and Xho I (3'). The open reading frame was derived from and the 3’ RE. Internal SalI sites may be present. The insert was GNG1100000 by the PCR. Insert size = 250 bp. sequenced and found to be identical with GB ACC#AF493875. Insert size = 223 bp. GNG120HN00 G-protein gamma 12 HA-tagged (N-terminus) N-terminal hemagglutinin tagged human G-protein gamma 12 GNG0900001 G-protein gamma 9 (pDNR-1r) subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI Human G-protein gamma 9 subunit (wild type) GNG0900000 (5') and Xho I (3'). The open reading frame was derived from cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GNG1200000 by the PCR. Insert size = 250 bp. SmaI site was destroyed (*). The insert canbe excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert GNG130HN00 G-protein gamma 13 HA tagged (N-terminus) was sequenced and found to be idential with GB N-terminal hemagglutinin tagged human G-protein gamma 13 ACC#AF493876. Insert size = 219 bp. Predicted amino acid subunit (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI sequence is 86% identical to bovine G- protein gamma 9 (5') and Xho I (3'). The open reading frame was derived from (U20085). GNG1300000 by the PCR. Insert size = 230 bp. GNG1000001 G-protein gamma 10 (pDNR-1r) Human G-protein gamma 10 subunit (wild type) GNG1000000 Gamma Wild-Type in pDNR-1r. Native G-protein cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The gamma subunits in pDNR-1r vector SmaI site was destroyed(* ). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was GNG0100001 G-protein gamma 1 (pDNR-1r) sequenced and found to be identical with GB ACC# AF493877. Human G-protein gamma subunit 1 (wild type) GNG0100000 Insert size = 220 bp. cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI GNG1100001 G-protein gamma 11 (pDNR-1r) and the 3’ RE. Internal SalI sites may be present. The insert was Human G-protein gamma 11 subunit (wild type) GNG1100000 sequenced and found to be identical with GB ACC# AF493869. cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The Insert size = 225 bp. SmaI site was destroyed(* ). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was GNG0200001 G-protein gamma 2 (pDNR-1r) sequenced and found to be identical with GB ACC# AF493878. Human G-protein gamma subunit 2 (wild type) GNG0200000 Insert size = 235 bp. cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI GNG1200001 G-protein gamma 12 (pDNR-1r) and the 3’ RE. Internal SalI sites may be present. The insert was Human G-protein gamma 12 subunit (wild type) GNG1200000 sequenced and found to be identical with GB ACC# AF493870. cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The Insert size = 220 bp. SmaI site was destroyed(* ). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was GNG0300001 G-protein gamma 3 (pDNR-1r) sequenced and found to be identical with GB ACC# AF493879. Human G-protein gamma 3 subunit (wild type) cloned into Insert size = 227 bp. pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ GNG1300001 G-protein gamma 13 (pDNR-1r) RE. Internal SalI sites may be present. The insert was Human G-protein gamma 13 subunit (wild type) GNG1300000

37 cDNA Resource Center

cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(* ). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical to GB ACC# AF493880. The predicted amino acid sequence is identical to that reported by Huang et al. (1999). Insert size = 213 bp.

38 cDNA Resource Center

G Protein Regulators

AGS proteins Activators of G protein signaling RCC1 Proteins Regulator of condensation 1 (RCC1) and guanine nucleotide exchange factor (GEF) for Ran proteins RGS Proteins Heterotrimeric G- protein GAPs; regulators of G- protein signaling (RGS) proteins RHO GDI Proteins Rho family guanine nucleotide dissociation inhibitor (GDI) proteins

AGS Proteins RGS0200000 RGS 2 (a.k.a: G0 to G1 switch regulatory 8, 24kD). GenBank Accession AGS0100000 Human activator of G-protein signaling AGS1 # AF493926: Human RGS2 (wild type) cloned into pcDNA3.1+ Human activator of G-protein signaling AGS1 (wild type) cloned (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The was amplified by the PCR from Image clone (43727). The insert open reading frame was amplified by the PCR from human was sequenced. Insert size = 640 bp. whole brain cDNA (Clontech). The insert was sequenced. Insert RGS020TN00 RGS 2 3xHA-tagged (N-terminus) size = 850 bp. RGS030TN00 RGS 3 3xHA-tagged (N-terminus) AGS030000R Rat activator of G-protein signaling AGS3 N-terminal 3X-hemagglutinin tagged human RGS3 (wild type) Rat activator of G-protein signaling AGS3 (wild type) cloned into cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open The open reading frame was derived from RGS0300000 by the reading frame was amplified by the PCR from rat whole brain PCR. Insert size = 1670 bp. cDNA. The insert was sequenced. Insert size = 1960 bp. RGS03T0000 RGS 3T AGS03L0000 Human Activator of G-protein Signaling AGS3-like Human RGS3T (wild type) cloned intopcDNA3.1+ (Invitrogen) at (Alternate Symbols: AGS3, GPSM1) GenBank Accession # KpnI (5') and XhoI (3'). The open reading frame was amplified by AY173053: Human activator of G-protein signaling AGS3 (wild the PCR from human whole brain cDNA (Clontech). The insert type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and XhoI was sequenced. Insert size = 630 bp. (3'). The open reading frame was amplified by the PCR from RGS03TTN00 RGS 3T 3xHA-tagged (N-terminus) human cDNA (Clontech). The insert was sequenced. Insert size = 1959 bp. RGS0400000 RGS 4 Human RGS4 (wild type) cloned into pcDNA3.1+ (Invitrogen) at AGS03S000R Rat activator of G-protein signaling AGS3 short KpnI (5') and XhoI (3'). The open reading frame was amplified by Rat activator of G-protein signaling AGS3 short isoform (wild the PCR from human whole brain cDNA (Clontech). The insert type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI was sequenced. Insert size = 620 bp. (3'). The open reading frame was amplified by the PCR from rat RGS040TN00 RGS 4 3xHA-tagged (N-terminus) whole brain cDNA. The insert was sequenced. Insert size = 510 bp. RGS0500000 RGS 5 Human RGS5 (wild type) cloned into pcDNA3.1+ (Invitrogen) at LGN0000000 AGS3-like (LGN) KpnI (5') and XhoI (3'). The open reading frame was amplified by Human LGN protein (wild type) G-protein signalling modulator 2 the PCR from Image clone (550112). The insert was sequenced. (AGS3-like, C. elegans) (GPSM2) cloned into pcDNA3.1+ Insert size = 550 bp. (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame RGS050TN00 RGS 5 3xHA-tagged (N-terminus) was amplified by the PCR from human whole brain cDNA. The insert was sequenced. Insert size = 2055 bp. RGS0700000 RGS 7 Human RGS7(wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was amplified by RCC1 Protein the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 1410 bp. RCC0100000 RCC1 Human regulator of chromosome condensation 1 RCC1 (wild RGS0710000 RGS 7 (S1) type) and guanine nucleotide exchange factor (GEF) for Ran Human RGS7(S1) (wild type, splice variant) cloned into proteins. Cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and pcDNA3.1+ (Invitrogen) at KpnI (5') and Xho I (3'). The open XhoI (3'). The open reading frame was amplified by the PCR reading frame was amplified by the PCR from human whole from human whole brain cDNA (clontech). The insert was brain cDNA (Clontech). The insert was sequenced and found to sequenced. Insert size = 1280 bp. be a splice variant of GB ACC# AF090116. When compared to AF090116 the following differences were noted: 1) deletion of bp 225-385 (residues 76-128); 2) A701->G (Q234R); 3) 78 bp RGS Proteins insertion between 1362-1363 (new 23 residue C-terminus). Insert size = 1330 bp. RGS0100000 RGS 1 RGS071TN00 RGS 7 (S1) 3xHA-tagged (N-terminus) Human RGS1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and NotI (3'). The open reading frame was amplified by RGS0720000 RGS 7 (S2) the PCR from Image clone (1519345). The insert was Human RGS7(S2) (wild type, splice variant) cloned into sequenced. Insert size = 600 bp. pcDNA3.1+ (Invitrogen) at KpnI (5') and Xho I (3'). The open RGS010TN00 RGS 1 3xHA-tagged (N-terminus) reading frame was amplified by the PCR from human whole

39 cDNA Resource Center

brain cDNA (Clontech). The insert was sequenced and found to following exeption (C690T) silent mutation: cloned into be a splice variant of GB ACC# AF090116. When compared to pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open AF090116 the following differences were noted: 1) A77->G reading frame was amplified by the PCR from human lung cDNA (K26R); 2) 78 bp insertion between 1362-1363 (new 23 residue (Clontech). Insert size = 708 bp. C-terminus). Insert size = 1490 bp. RGS072TN00 RGS 7 (S2) 3xHA-tagged (N-terminus) RGS1900000 RGS 19 (a.k.a: G alpha interacting protein, guanine nucleotide binding RGS0800000 RGS 8 protein alpha inhibiting activity polypeptide interacting protein). Human RGS8 (wild type) cloned into pcDNA3.1+ (Invitrogen) at GenBank Accession # AF493939: Human RGS protein KpnI (5') and XhoI (3'). The open reading frame was amplified by GAIP/RGS19 (wild type) cloned into pcDNA3.1+ (Invitrogen) at the PCR from human whole brain cDNA. The insert was KpnI (5') and XhoI (3'). The open reading frame was amplified by sequenced and the predicted amino acid sequence found to be the PCR from IMAGE clone (33393). The insert was sequenced. 99% identical to rat RGS8. Insert size = 550 bp. Insert size = 660 bp. RGS080TN00 RGS 8 3xHA-tagged (N-terminus) RGS190TN00 RGS 19 3xHA-tagged (N-terminus)

RGS1000000 RGS 10 RGSZ100000 RGS Z1 Human RGS14 short (wild type) cloned into pcDNA3.1+ (a.k.a: Regulator of Gz-selective protein signaling). GenBank (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame Accession # AF493940: Human RGSZ1/RGS20 (wild type) was amplified by the PCR from Image clone (2275604). The cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). insert was sequenced. Insert size = 610 bp. The open reading frame was amplified by the PCR from Image RGS100TN00 RGS 10 3xHA-tagged (N-terminus) clone (1690771). The insert was sequenced. Insert size = 660 bp. RGS1300000 RGS 13 RGSZ10TN00 RGS Z1 3xHA-tagged (N-terminus) Human RGS13 (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was amplified by the PCR from Image clone (239446). The insert was sequenced. Insert size = 485 bp. RHO GDI Proteins RGS130TN00 RGS 13 3xHA-tagged (N-terminus) RHI0A00000 Rho GDI alpha RGS14S0000 RGS 14 short Human Rho GDP dissociation inhibitor (Rho GDI alpha) (wild Human RGS14 short (wild type) cloned into pcDNA3.1+ type) cloned into pcDNA3.1+ (Invitrogen) at EcoR I (5') and Xho I (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame (3'). The open reading frame was amplified by the PCR was amplified by the PCR from Image clone (2275604). The fromHuman whole brain cDNA (Clontech). The insert was insert was sequenced. Insert size = 610 bp. sequenced. Insert size = 625 bp RGS14STN00 RGS 14 short 3xHA-tagged(N-terminus) RHI0B00000 Rho GDI beta RGS1600000 RGS 16 Human Rho GDP dissociation inhibitor (Rho GDI beta) (wild Human RGS16 (wild type) cloned into pcDNA3.1+ (Invitrogen) at type) cloned into pcDNA3.1+ (Invitrogen) at EcoR I (5') and Xho I KpnI (5') and XhoI (3'). The open reading frame was amplified by (3'). The open reading frame was amplified by the PCR the PCR from Image clone (2030792). The insert was fromHuman whole brain cDNA (Clontech). The insert was sequenced. Insert size = 615 bp. sequenced. Insert size = 615 bp. RGS160TN00 RGS 16 3xHA-tagged (N-terminus) RHI0G00000 Rho GDI gamma RGS1700000 RGS 17 Human Rho GDP dissociation inhibitor (Rho GDI gamma) (wild Human RGS17 (wild type) cloned into pcDNA3.1+ (Invitrogen) at type) cloned into pcDNA3.1+ (Invitrogen) at EcoR I (5') and Xho I KpnI (5') and XbaI (3'). The open reading frame was amplified by (3'). The open reading frame was amplified by the PCR the PCR fromHuman whole brain cDNA (Clontech). The insert fromHuman whole brain cDNA (Clontech). The insert was was sequenced. Insert size = 470 bp. sequenced. Insert size = 690 bp. RGS170TN00 RGS 17 3xHA-tagged (N-terminus)

RGS1800000 RGS 18 RGS18 identical to Genbank ACC# NM_130782 with the

40 cDNA Resource Center

Small G-proteins

Arf family Rab family Ran family Ras family Ras-like Rho family

Arf Family Wild-Type Native ADP ribosylating factors (ARF) and ARF-like (ARL) proteins Wild -Type in pDNR-1r Native ADP ribosylating factors (ARF) and ARF-like (ARL) proteins in pDNR-1r vector

Arf Wild-Type frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 570 bp. ARF0100000 ARF1 GenBank Accession # AF493881: Human ADP-ribosylation ARL0300000 ARL3 factor protein ARF1 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF493889: Human ADP-ribosylation (Invitrogen) at BamH I (5') and Xho I (3'). The open reading factor-like protein ARL3 (wild type) cloned into pcDNA3.1+ frame was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamH I (5') and Xho I (3'). The open reading (Clontech). The insert was sequenced. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 560 bp. ARF0300000 ARF3 GenBank Accession # AF493882: Human ADP-ribosylation ARL0400000 ARL4 factor protein ARF3 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF493890: Human ADP-ribosylation (Invitrogen) at BamH I (5') and Xho I (3'). The open reading factor-like protein ARL4 (wild type) cloned into pcDNA3.1+ frame was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamH I (5') and Xho I (3'). The open reading (Clontech). The insert was sequenced. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 610 bp. ARF0400000 ARF4 GenBank Accession # AF493883: Human ADP-ribosylation ARL0500000 ARL5 factor protein ARF4 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF493891: Human ADP-ribosylation (Invitrogen) at BamH I (5') and Xho I (3'). The open reading factor-like protein ARL5 (wild type) cloned into pcDNA3.1+ frame was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamH I (5') and Xho I (3'). The open reading (Clontech). The insert was sequenced. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 550 bp. ARF0500000 ARF5 GenBank Accession # AF493884: Human ADP-ribosylation ARL0700000 ARL7 factor protein ARF5 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF493892: Human ADP-ribosylation (Invitrogen) at BamH I (5') and Xho I (3'). The open reading factor-like protein ARL7 (wild type) cloned into pcDNA3.1+ frame was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamH I (5') and Xho I (3'). The open reading (Clontech). The insert was sequenced. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 590 bp. ARF0600000 ARF6 GenBank Accession # AF493885: Human ADP-ribosylation factor protein ARF6 (wild type) cloned into pcDNA3.1+ Arf Wild-Type in pDNR-1r (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA ARF0100001 ARF1 (pDNR-1r) (Clontech). The insert was sequenced. Insert size = 545 bp. Human ADP-ribosylation factor protein ARF1 (wild type) ARF0100000 cloned into pDNR-1r (Clontech) at SmaI (5') and ARF0P00000 ARF protein XhoI (3'). The SmaI site was destroyed(* ). The insert can be GenBank Accession # AF493886: Human ADP-ribosylation excised with SalI and the 3’ RE. Internal SalI sites may be factor-like protein ARF protein (wild type) cloned into pcDNA3.1+ present. The insert was sequenced and found to be identical with (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GB ACC # AF493881. Insert size = 560 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 410 bp. ARF0400001 ARF4 (pDNR-1r) Human ADP-ribosylation factor protein ARF4 (wild type) ARL0100000 ARL1 ARF0400000 cloned into pDNR-1r (Clontech) at SmaI (5') and GenBank Accession # AF493887: Human ADP-ribosylation XhoI (3'). The SmaI site was destroyed(* ). The insert can be factor-like protein ARL1 (wild type) cloned into pcDNA3.1+ excised with SalI and the 3’ RE. Internal SalI sites may be (Invitrogen) at BamH I (5') and Xho I (3'). The open reading present. The insert was sequenced and found to be identical with frame was amplified by the PCR from human whole brain cDNA GB ACC # AF493883. Insert size = 560 bp. (Clontech). The insert was sequenced. Insert size = 560 bp. ARF0500001 ARF5 (pDNR-1r) ARL0200000 ARL2 Human ADP-ribosylation factor protein ARF5 (wild type) GenBank Accession # AF493888: Human ADP-ribosylation ARF0500000 cloned into pDNR-1r (Clontech) at SmaI (5') and factor-like protein ARL2 (wild type) cloned into pcDNA3.1+ XhoI (3'). The SmaI site was destroyed(* ). The insert can be (Invitrogen) at BamH I (5') and Xho I (3'). The open reading excised with SalI and the 3’ RE. Internal SalI sites may be

41 cDNA Resource Center

present. The insert was sequenced and found to be identical with XhoI (3'). The SmaI site was destroyed(*). The insert can be GB ACC # AF493884. Insert size = 560 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with ARF0600001 ARF6 (pDNR-1r) GB ACC # AF493889. Insert size = 560 bp. Human ADP-ribosylation factor protein ARF6 (wild type) ARF0600000 cloned into pDNR-1r (Clontech) at SmaI (5') and ARL0400001 ARL4 (pDNR-1r) XhoI (3'). The SmaI site was destroyed(* ). The insert can be Human ADP-ribosylation factor-like protein ARL4 (wild type) excised with SalI and the 3’ RE. Internal SalI sites may be ARL0400000 cloned into pDNR-1r (Clontech) at SmaI (5') and present. The insert was sequenced and found to be identical with XhoI (3'). The SmaI site was destroyed(*). The insert can be GB ACC # AF493885. Insert size = 545 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with ARL0100001 ARL1 (pDNR-1r) GB ACC # AF493890 with the following exception:A105- Human ADP-ribosylation factor-like protein ARL1 (wild type) >T(silent). Insert size = 610 bp. ARL0100000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be ARL0500001 ARL5 (pDNR-1r) excised with SalI and the 3’ RE. Internal SalI sites may be Human ADP-ribosylation factor-like protein ARL5 (wild type) present. The insert was sequenced and found to be identical with ARL0500000 cloned into pDNR-1r (Clontech) at SmaI (5') and GB ACC # AF493887. Insert size = 560 bp. XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be ARL0200001 ARL2 (pDNR-1r) present. The insert was sequenced and found to be identical with Human ADP-ribosylation factor-like protein ARL2 (wild type) GB ACC # AF493891. Insert size = 550 bp. ARL0200000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be ARL0700001 ARL7 (pDNR-1r) excised with SalI and the 3’ RE. Internal SalI sites may be Human ADP-ribosylation factor-like protein ARL7 (wild type) present. The insert was sequenced and found to be identical with ARL0700000 cloned into pDNR-1r (Clontech) at SmaI (5') and GB ACC # AF493888. Insert size = 570 bp. XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be ARL0300001 ARL3 (pDNR-1r) present. The insert was sequenced and found to be identical with Human ADP-ribosylation factor-like protein ARL3 (wild type) GB ACC # AF493892. Insert size = 590 bp. ARL0300000 cloned into pDNR-1r (Clontech) at SmaI (5') and

Rab family

Wild-Type Native Rab proteins Wild-Type in pDNR-1r Native Rab proteins in pDNR-1r vector GTP binding protein RAB3D (wild type) cloned into pcDNA3.1+ Rab Wild-Type (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA RAB01A0000 RAB 1A (Clontech). The insert was sequenced. Insert size = 670 bp. GenBank Accession # AF498929: Human RAB family small GTP binding protein RAB1A (wild type) cloned into pcDNA3.1+ RAB04A0000 RAB 4A (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GenBank Accession # AF498934: Human RAB family small was amplified by the PCR from human whole brain cDNA GTP binding protein RAB4A (wild type) cloned into pcDNA3.1+ (Clontech). The insert was sequenced. Insert size = 625 bp. (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA RAB02A0000 RAB 2A (Clontech). The insert was sequenced. Insert size = 670 bp. GenBank Accession # AF498930: Human RAB family small GTP binding protein RAB2A (wild type) cloned into pcDNA3.1+ RAB04B0000 RAB 4B (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GenBank Accession # AF498935: Human RAB family small was amplified by the PCR from human whole brain cDNA GTP binding protein RAB4B (wild type) cloned into pcDNA3.1+ (Clontech). The insert was sequenced. Insert size = 650 bp. (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA RAB03A0000 RAB 3A (Clontech). The insert was sequenced. Insert size = 650 bp. GenBank Accession # AF498931: Human RAB family small GTP binding protein RAB3A (wild type) cloned into pcDNA3.1+ RAB05A0000 RAB 5A (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GenBank Accession # AF498936: Human RAB family small was amplified by the PCR from human whole brain cDNA GTP binding protein RAB5A (wild type) cloned into pcDNA3.1+ (clontech). The insert was sequenced. Insert size = 675 bp. (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA RAB03B0000 RAB 3B (Clontech). The insert was sequenced. Insert size = 655 bp. GenBank Accession # AF498932: Human RAB family small GTP binding protein RAB3B (wild type) cloned into pcDNA3.1+ RAB05B0000 RAB 5B (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GenBank Accession # AF498937: Human RAB family small was amplified by the PCR from human whole brain cDNA GTP binding protein RAB5B (wild type) cloned into pcDNA3.1+ (Clontech). The insert was sequenced. Insert size = 675 bp. (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA RAB03D0000 RAB 3D (Clontech). The insert was sequenced. Insert size = 670 bp. GenBank Accession # AF498933: Human RAB family small

42 cDNA Resource Center

RAB05C0000 RAB 5C RAB1300000 RAB 13 GenBank Accession # AF498938: Human RAB family small GenBank Accession # AF498948: Human RAB family small GTP binding protein RAB5C (wild type) cloned into pcDNA3.1+ GTP binding protein RAB13 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Invitrogen) at BamHI (5') and NotI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 660 bp. (Clontech). The insert was sequenced. Insert size = 620 bp.

RAB06A0000 RAB 6A RAB1400000 RAB 14 GenBank Accession # AF498939: Human RAB family small GenBank Accession # AF498949: Human RAB family small GTP binding protein RAB6A (wild type) cloned into pcDNA3.1+ GTP binding protein RAB14 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 635 bp. (Clontech). The insert was sequenced. Insert size = 660 bp.

RAB06B0000 RAB 6B RAB1800000 RAB 18 GenBank Accession # AF498940: Human RAB family small GenBank Accession # AF498950: Human RAB family small GTP binding protein RAB6B (wild type) cloned into pcDNA3.1+ GTP binding protein RAB18 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 630 bp. (Clontech). The insert was sequenced. Insert size = 630 bp.

RAB06C0000 RAB 6C RAB2300000 RAB 23 GenBank Accession # AF498941: Human RAB family small GenBank Accession # AF498951: Human RAB family small GTP binding protein RAB6C (wild type) cloned into pcDNA3.1+ GTP binding protein RAB23 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Invitrogen) at HindIII (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 630 bp. (Clontech). The insert was sequenced. Insert size = 730 bp.

RAB0700000 RAB 7 RAB2600000 RAB 26 GenBank Accession # AF498942: Human RAB family small GenBank Accession # AF498952: Human RAB family small GTP binding protein RAB7 (wild type) cloned into pcDNA3.1+ GTP binding protein RAB26 (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 630 bp. (clontech). The insert was sequenced and found to be identical with GB ACC#NM_014353. Insert size = 580 bp. RAB0800000 RAB 8 GenBank Accession # AF498943: Human RAB family small RAB27A0000 RAB 27a GTP binding protein RAB8 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498953: Human RAB family small (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GTP binding protein RAB27a (wild type) cloned into pcDNA3.1+ was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Clontech). The insert was sequenced. Insert size = 630 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 670 bp. RAB0900000 RAB 9 GenBank Accession # AF498944: Human RAB family small RAB27B0000 RAB 27B GTP binding protein RAB9 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498954: Human RAB family small (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GTP binding protein RAB27B (wild type) cloned into pcDNA3.1+ was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Clontech). The insert was sequenced. Insert size = 615 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 660 bp. RAB1000000 RAB 10 GenBank Accession # AF498945: Human RAB family small RAB2800000 RAB 28 GTP binding protein RAB10 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498955: Human RAB family small (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GTP binding protein RAB28 (wild type) cloned into pcDNA3.1+ was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Clontech). The insert was sequenced. Insert size = 610 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 580 bp. RAB11A0000 RAB 11A GenBank Accession # AF498946: Human RAB family small RAB3000000 RAB 30 GTP binding protein RAB11A (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498956: Human RAB family small (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GTP binding protein RAB30 (wild type) cloned into pcDNA3.1+ was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Clontech). The insert was sequenced. Insert size = 660 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 630 bp. RAB11B0000 RAB 11B GenBank Accession # AF498947: Human RAB family small RAB3100000 RAB 31 GTP binding protein RAB11B (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498957: Human RAB family small (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame GTP binding protein RAB31 (wild type) cloned into pcDNA3.1+ was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame (Clontech). The insert was sequenced. Insert size = 675 bp. was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 590 bp.

43 cDNA Resource Center

RAB3200000 RAB 32 RAB04B0001 Rab 4B (pDNR-1r) GenBank Accession # AF498958: Human RAB family small Human RAB family small GTP binding protein RAB4B (wild type) GTP binding protein RAB32 (wild type) cloned into pcDNA3.1+ RAB04B0000 cloned into pDNR-1r (Clontech) at SmaI (5') and (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame XhoI (3'). The SmaI site was destroyed (*). The insert can be was amplified by the PCR from human whole brain cDNA excised with SalI and the 3’ RE. Internal SalI sites may be (Clontech). The insert was sequenced. Insert size = 690 bp. present. The insert was sequenced and found to be identical with GB ACC # AF498935. Insert size = 650 bp. RAB33A0000 RAB 33A GenBank Accession # AF498959: Human RAB family small RAB05A0001 Rab 5A (pDNR-1r) GTP binding protein RAB33A (wild type) cloned into pcDNA3.1+ Human RAB family small GTP binding protein RAB5A (wild type) (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame RAB05A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and was amplified by the PCR from human whole brain cDNA XhoI (3'). The SmaI site was destroyed (*). The insert can be (Clontech). The insert was sequenced. Insert size = 720 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with RAB3500000 RAB 35 GB ACC # AF498936. Insert size = 655 bp. GenBank Accession # AF498960: Human RAB family small GTP binding protein RAB35 (wild type) cloned into pcDNA3.1+ RAB05B0001 Rab 5B (pDNR-1r) (Invitrogen) at BamHI (5') and XhoI (3'). The open reading frame Human RAB family small GTP binding protein RAB5B (wild type) was amplified by the PCR from human whole brain cDNA RAB05B0000 cloned into pDNR-1r (Clontech) at SmaI (5') and (Clontech). The insert was sequenced. Insert size = 615 bp. XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be

present. The insert was sequenced and found to be identical with Rab Wild-Type in pDNR-1r GB ACC # AF498937. Insert size = 670 bp.

RAB01A0001 Rab 1A (pDNR-1r) Human RAB family small GTP binding protein RAB1A (wild type) RAB05C0001 Rab 5C (pDNR-1r) RAB01A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and Human RAB family small GTP binding protein RAB5C (wild type) XhoI (3'). The SmaI site was destroyed (*). The insert can be RAB05C0000 cloned into pDNR-1r (Clontech) at SmaI (5') and excised with SalI and the 3’ RE. Internal SalI sites may be XhoI (3'). The SmaI site was destroyed (*). The insert can be present. The insert was sequenced and found to be identical with excised with SalI and the 3’ RE. Internal SalI sites may be GB ACC # AF498929. Insert size = 625 bp. present. The insert was sequenced and found to be identical with GB ACC # AF498938. Insert size = 660 bp. RAB02A0001 Rab 2A (pDNR-1r) Human RAB family small GTP binding protein RAB2A (wild type) RAB06A0001 Rab 6A (pDNR-1r) RAB02A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and Human RAB family small GTP binding protein RAB6A (wild type) XhoI (3'). The SmaI site was destroyed (*). The insert can be RAB06A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and excised with SalI and the 3’ RE. Internal SalI sites may be XhoI (3'). The SmaI site was destroyed (*). The insert can be present. The insert was sequenced and found to be identical with excised with SalI and the 3’ RE. Internal SalI sites may be GB ACC # AF498930. Insert size = 650 bp. present. The insert was sequenced and found to be identical with GB ACC # AF498939. Insert size = 635 bp. RAB03A0001 Rab 3A (pDNR-1r) Human RAB family small GTP binding protein RAB3A (wild type) RAB06B0001 Rab 6B (pDNR-1r) RAB03A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and Human RAB family small GTP binding protein RAB6B (wild type) XhoI (3'). The SmaI site was destroyed (*). The insert can be RAB06B0000 cloned into pDNR-1r (Clontech) at SmaI (5') and excised with SalI and the 3’ RE. Internal SalI sites may be XhoI (3'). The SmaI site was destroyed (*). The insert can be present. The insert was sequenced and found to be identical with excised with SalI and the 3’ RE. Internal SalI sites may be GB ACC # AF498931. Insert size = 675 bp. present. The insert was sequenced and found to be identical with GB ACC # AF498940. Insert size = 630 bp. RAB03B0001 Rab 3B (pDNR-1r) Human RAB family small GTP binding protein RAB3B (wild type) RAB06C0001 Rab 6C (pDNR-1r) RAB03B0000 cloned into pDNR-1r (Clontech) at SmaI (5') and Human RAB family small GTP binding protein RAB6C (wild type) XhoI (3'). The SmaI site was destroyed (*). The insert can be RAB06C0000 cloned into pDNR-1r (Clontech) at SmaI (5') and excised with SalI and the 3’ RE. Internal SalI sites may be XhoI (3'). The SmaI site was destroyed (*). The insert can be present. The insert was sequenced and found to be identical with excised with SalI and the 3’ RE. Internal SalI sites may be GB ACC # AF498932. Insert size = 675 bp. present. The insert was sequenced and found to be identical with GB ACC # AF498941. Insert size = 630 bp.

RAB03D0001 Rab 3D (pDNR-1r) RAB0700001 Rab 7 (pDNR-1r) Human RAB family small GTP binding protein RAB3D (wild type) Human RAB family small GTP binding protein RAB7 (wild type) RAB03D0000 cloned into pDNR-1r (Clontech) at SmaI (5') and RAB0700000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with present. The insert was sequenced and found to be identical with GB ACC # AF498933. Insert size = 670 bp. GB ACC # AF498942. Insert size = 630 bp.

RAB04A0001 Rab 4A (pDNR-1r) RAB0800001 Rab 8 (pDNR-1r) Human RAB family small GTP binding protein RAB4A (wild type) Human RAB family small GTP binding protein RAB8 (wild type) RAB04A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and RAB0800000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with present. The insert was sequenced and found to be identical with GB ACC # AF498934. Insert size = 670 bp. GB ACC # AF498943. Insert size = 630 bp.

44 cDNA Resource Center

RAB0900001 Rab 9 (pDNR-1r) RAB2300001 Rab 23 (pDNR-1r) Human RAB family small GTP binding protein RAB9 (wild type) Human RAB family small GTP binding protein RAB23 (wild type) RAB0900000 cloned into pDNR-1r (Clontech) at SmaI (5') and RAB2300000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with present. The insert was sequenced and found to be identical with GB ACC # AF498944. Insert size = 615 bp. GB ACC # AF498951. Insert size = 730 bp.

RAB1000001 Rab 10 (pDNR-1r) RAB2600001 Rab 26 (pDNR-1r) Human RAB family small GTP binding protein RAB10 (wild type) Human RAB family small GTP binding protein RAB26 (wild type) RAB1000000 cloned into pDNR-1r (Clontech) at SmaI (5') and RAB2600000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with present. The insert was sequenced and found to be identical with GB ACC # AF498945. Insert size = 610 bp. GB ACC # AF498952. Insert size = 580 bp.

RAB11A0001 Rab 11A (pDNR-1r) RAB27A0001 Rab 27A (pDNR-1r) Human RAB family small GTP binding protein RAB11A (wild Human RAB family small GTP binding protein RAB27a (wild type) RAB11A0000 cloned into pDNR-1r (Clontech) at SmaI (5') type) RAB27A0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be and XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be excised with SalI and the 3’ RE. Internal SalI sites present. The insert was sequenced and found to be identical with may be present. The insert was sequenced and GB ACC # AF498946. Insert size = 660 bp. found to be identical with GB ACC # AF498954. RAB11B0001 Rab 11B (pDNR-1r) Insert size = 670 bp. Human RAB family small GTP binding protein RAB11B (wild type) RAB11B0000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC # AF498947. Insert size = 675 bp.

Ran family

Wild-Type Native RAN family proteins 3xHA-Tagged Native RAN family proteins with an N-terminal hemagglutinin tag in triplicate

Ran Wild-Type Ran 3xHA-Tagged

RAN0000000 RAN RAN000TN00 RAN 3xHA-tagged(N-terminus) GenBank Accession # AF501887: Human RAN family small N-terminal 3X-hemagglutinin tagged human RAN family small GTP binding protein RAN (wild type) cloned into pcDNA3.1+ GTP binding protein RAN (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame frame was amplified by the PCR from human whole brain was derived from RAN0000000 by the PCR. Insert size = 750 cDNA (Clontech). The insert was sequenced. Insert size = 660 bp. bp.

45 cDNA Resource Center

Ras Family

Wild-Type Native Ras family proteins pDNR-1r Native Ras family proteins in pDNR-1r vector GV Mutants Ras family proteins with a constitutively active phenotype due to a glycine (G) to valine (V) mutation, which eliminates GTPase activity 3xHA GV Ras family proteins with a constitutively active phenotype and an N-terminal hemagglutinin tag in triplicate SN Mutants Ras family proteins with a dominant negative phenotype due to a serine (S) to asparagine (N) mutation 3xHA SN Ras family proteins with a dominant negative phenotype and an N-terminal hemagglutinin tag in triplicate 3xHA-Tagged Native Ras family proteins with an N-terminal hemagglutinin tag in triplicate 2XMYC GV Ras family proteins with a constitutively active phenotype and an N-terminal MYC tag in duplicate 2XMYC SN Ras family proteins with a dominant negative phenotype and an N-terminal MYC tag in duplicate 2XMYC-tagged Native Ras family proteins with an N-terminal MYC tag in duplicate

Ras Family Wild-Type frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 580 bp. RAP1A00000 RAP1A GenBank Accession # AF493912: Human Ras family small GTP RASR000000 R-Ras binding protein RAP1A (wild type) cloned into pcDNA3.1+ GenBank Accession # AF493920: Human Ras family small GTP (Invitrogen) at BamH I (5') and Xho I (3'). The open reading binding protein R-Ras (wild type) cloned into pcDNA3.1+ frame was amplified by the PCR from human whole brain cDNA (Invitrogen) at BamH I (5') and Xho I (3'). The open reading (Clontech). The insert was sequenced. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 660 bp. RAP1B00000 RAP1B GenBank Accession # AF493913: Human Ras family small GTP Ras Family Wild-Type in pDNR-1r binding protein RAP1B (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamH I (5') and Xho I (3'). The open reading RAP1A00001 RAP 1A (pDNR-1R) frame was amplified by the PCR from human whole brain cDNA Human Ras family small GTP binding protein RAP1A (wild type) (Clontech). The insert was sequenced. Insert size = 560 bp. RAP1A00000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be RAP2A00000 RAP2A excised with SalI and the 3’ RE. Internal SalI sites may be GenBank Accession # AF493914: Human Ras family small GTP present. The insert was sequenced and found to be identical with binding protein RAP2A (wild type) cloned into pcDNA3.1+ GB ACC # AF493912. Insert size = 560 bp. (Invitrogen) at BamH I (5') and Xho I (3'). The open reading frame was amplified by the PCR from Image clone (2407404). RAP1B00001 RAP1B (pDNR-1r) The insert was sequenced. Insert size = 560 bp. Human Ras family small GTP binding protein RAP1B (wild type) RAP1B00000 cloned into pDNR-1r (Clontech) at SmaI (5') and RAP2B00000 RAP2B XhoI (3'). The SmaI site was destroyed(*). The insert can be GenBank Accession # AF493915: Human Ras family small GTP excised with SalI and the 3’ RE. Internal SalI sites may be binding protein RAP2A (wild type) cloned into pcDNA3.1+ present. The insert was sequenced and found to be identical with (Invitrogen) at BamH I (5') and Xho I (3'). The open reading GB ACC # AF493913. Insert size = 560 bp. frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 560 bp. RAP2A00001 RAP2A (pDNR-1r) Human Ras family small GTP binding protein RAP2A (wild type) RASH000000 H-Ras RAP2A00000 cloned into pDNR-1r (Clontech) at SmaI (5') and GenBank Accession # AF493916: Human Ras family small GTP XhoI (3'). The SmaI site was destroyed(*). The insert can be binding protein H-Ras (wild type) cloned into pcDNA3.1+ excised with SalI and the 3’ RE. Internal SalI sites may be (Invitrogen) at BamH I (5') and Xho I (3'). The open reading present. The insert was sequenced and found to be identical with frame was amplified by the PCR from human whole brain cDNA GB ACC # AF493914. Insert size = 560 bp. (Clontech). The insert was sequenced. Insert size = 570 bp. RAP2B00001 RAP2B (pDNR-1r) RASK200000 K-Ras2 Human Ras family small GTP binding protein RAP2B (wild type) GenBank Accession # AF493917: Human Ras family small GTP RAP2B00000 cloned into pDNR-1r (Clontech) at SmaI (5') and binding protein K-Ras2 (wild type) cloned into pcDNA3.1+ XhoI (3'). The SmaI site was destroyed(*). The insert can be (Invitrogen) at BamH I (5') and Xho I (3'). The open reading excised with SalI and the 3’ RE. Internal SalI sites may be frame was amplified by the PCR from human whole brain cDNA present. The insert was sequenced and found to be identical with (Clontech). The insert was sequenced. Insert size = 570 bp. GB ACC # AF493915. Insert size = 560 bp.

RASM000000 M-Ras RASH000001 H-Ras (pDNR-1r) GenBank Accession # AF493918: Human Ras family small GTP Human Ras family small GTP binding protein H-Ras (wild type) binding protein M-Ras (wild type) cloned into pcDNA3.1+ RASH000000 cloned into pDNR-1r (Clontech) at SmaI (5') and (Invitrogen) at BamH I (5') and Xho I (3'). The open reading XhoI (3'). The SmaI site was destroyed(*). The insert can be frame was amplified by the PCR from human whole brain cDNA excised with SalI and the 3’ RE. Internal SalI sites may be (Clontech). The insert was sequenced. Insert size = 630 bp. present. The insert was sequenced and found to be identical with GB ACC # AF493916. Insert size = 570 bp. RASN000000 N-Ras GenBank Accession # AF493919: Human Ras family small GTP RASK200001 K-Ras2 (pDNR-1r) binding protein N-Ras (wild type) cloned into pcDNA3.1+ Human Ras family small GTP binding protein K-Ras2 (wild type) (Invitrogen) at BamH I (5') and Xho I (3'). The open reading RASK200000 cloned into pDNR-1r (Clontech) at SmaI (5') and

46 cDNA Resource Center

XhoI (3'). The SmaI site was destroyed(*). The insert can be GTP binding protein N-Ras (RASN000000) via the Quickchange excised with SalI and the 3’ RE. Internal SalI sites may be mutagenesis kit (Stratagene). The mutation confers a present. The insert was sequenced and found to be identical with constitutively active phenotype. Insert size = 580 bp. GB ACC # AF493917. Insert size = 570 bp. RASR0000C0 R-Ras G38V RASM000001 M-Ras (pDNR-1r) The G38V mutation was introduced into the Ras family small Human Ras family small GTP binding protein M-Ras (wild type) GTP binding protein R-Ras (RASR000000) via the Quickchange RASM000000 cloned into pDNR-1r (Clontech) at SmaI (5') and mutagenesis kit (Stratagene). The mutation putatively confers a XhoI (3'). The SmaI site was destroyed(*). The insert can be constitutively active phenotype. Insert size = 660 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC # AF493918. Insert size = 630 bp. Ras GV Mutants 3xHA Tagged

RASN000001 N-Ras (pDNR-1r) RAP1A0TNC0 RAP1A G12V 3XHA-tagged (N-terminus) Human Ras family small GTP binding protein N-Ras (wild type) N-terminal 3XHemagglutinin-tagged human Ras family small RASN000000 cloned into pDNR-1r (Clontech) at SmaI (5') and GTP binding protein RAP1A G12V (constitutively active) cloned XhoI (3'). The SmaI site was destroyed(*). The insert can be into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V excised with SalI and the 3’ RE. Internal SalI sites may be mutation was introduced into the Ras family small GTP binding present. The insert was sequenced and found to be identical with protein Rap1A (RAP1A00000) via the Quickchange mutagenesis GB ACC # AF493919. Insert size = 580 bp. kit (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 640 bp. RASR000001 R-Ras (pDNR-1r) Human Ras family small GTP binding protein R-Ras (wild type) RAP1B0TNC0 RAP1B G12V 3XHA-tagged (N-terminus) RASR000000 cloned into pDNR-1r (Clontech) at SmaI (5') and N-terminal 3Xhemagglutinin-tagged human Ras family small XhoI (3'). The SmaI site was destroyed(*). The insert can be GTP binding protein RAP1B G12V (constitutively active) cloned excised with SalI and the 3’ RE. Internal SalI sites may be into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V present. The insert was sequenced and found to be identical with mutation was introduced into the Ras family small GTP binding GB ACC # AF493920. Insert size = 660 bp. protein Rap1B (RAP1B00000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 640 bp. Ras GV Mutants RASH00TNC0 H-Ras G12V 3xHA-tagged(N-terminus) RAP1A000C0 RAP1A G12V N-terminal 3x-hemagglutinin tagged human Ras family small Human Ras family small GTP binding protein Rap1A G12V GTP binding protein H-Ras (G12V mutant) cloned into (constitutively active) cloned into pcDNA3.1+ (Invitrogen) at pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V BamHI (5') and XhoI (3'). The G12V mutation was introduced mutation was introduced into H-Ras 3X HA tagged (N- into the Ras family small GTP binding protein Rap1A terminus)(RASH00TN00) via the Quickchange mutagenesis kit (RAP1A00000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 670 bp. phenotype. Insert size = 560 bp. RASK20TNC0 K-Ras2 G12V 3xHA-tagged(N-terminus) RAP1B000C0 RAP1B G12V N-terminal 3x-hemagglutinin tagged human Ras family small Human Ras family small GTP binding protein Rap1B G12V GTP binding protein K-Ras2 (G12V mutant) cloned into (constitutively active) cloned into pcDNA3.1+ (Invitrogen) at pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V BamHI (5') and XhoI (3'). The G12V mutation was introduced mutation was introduced into K-Ras2 3X HA tagged (N- into the Ras family small GTP binding protein Rap1B terminus)(RASK20TN00) via the Quickchange mutagenesis kit (RAP1B00000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 670 bp. phenotype. Insert size = 560 bp. RASM00TNC0 M-Ras G22V 3xHA-tagged(N-terminus) RASH0000C0 H-Ras G12V N-terminal 3x-hemagglutinin tagged human Ras family small The G12V mutation was introduced into the Ras family small GTP binding protein M-Ras (G22V mutant) cloned into GTP binding protein H-Ras (RASH000000) via the Quickchange pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G22V mutagenesis kit (Stratagene). The mutation confers a mutation was introduced into M-Ras 3X HA tagged (N- constitutively active phenotype. Insert size = 570 bp. terminus)(RASM00TN00) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active RASK2000C0 K-Ras2 G12V phenotype. Insert size = 730 bp. The G12V mutation was introduced into the Ras family small GTP binding protein K-Ras2 (RASK200000) via the RASN00TNC0 N-Ras G12V 3xHA-tagged(N-terminus) Quickchange mutagenesis kit (Stratagene). The mutation N-terminal 3x-hemagglutinin tagged human Ras family small confers a constitutively active phenotype. Insert size = 570 bp. GTP binding protein N-Ras (G12V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V RASM0000C0 M-Ras G22V mutation was introduced into N-Ras 3X HA tagged (N- The G22V mutation was introduced into the Ras family small terminus)(RASN00TN00) via the Quickchange mutagenesis kit GTP binding protein M-Ras (RASM000000) via the (Stratagene). The mutation confers a constitutively active Quickchange mutagenesis kit (Stratagene). The mutation phenotype. Insert size = 680 bp. confers a constitutively active phenotype. Insert size = 630 bp. RASR00TNC0 R-Ras G38V 3xHA-tagged(N-terminus) RASN0000C0 N-Ras G12V N-terminal 3x-hemagglutinin tagged human Ras family small The G12V mutation was introduced into the Ras family small GTP binding protein R-Ras (G38V mutant) cloned into

47 cDNA Resource Center

pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G38V GTP binding protein RAP1B S17N (dominant negative) cloned mutation was introduced into R-Ras 3X HA tagged (N- into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N terminus)(RASR00TN00) via the Quickchange mutagenesis kit mutation was introduced into the Ras family small GTP binding (Stratagene). The mutation confers a constitutively active protein Rap1B (RAP1B00000) via the Quickchange mutagenesis phenotype. Insert size = 760 bp. kit (Stratagene). The mutation confers a dominant negative phenotype. Insert size = 640 bp.

Ras Family SN Mutants RASH00TND0 H-Ras S17N 3xHA-tagged(N-terminus) N-terminal 3x-hemagglutinin tagged human Ras family small RAP1A000D0 RAP1A S17N GTP binding protein H-Ras (S17N mutant) cloned into Human Ras family small GTP binding protein RAP1A S17N pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N (dominant negative) cloned into pcDNA3.1+ (Invitrogen) at mutation was introduced into H-Ras 3X HA tagged (N- BamHI (5') and XhoI (3'). The S17N mutation was introduced into terminus)(RASH00TN00) via the Quickchange mutagenesis kit the Ras family small GTP binding protein Rap1A (RAP1A00000) (Stratagene). The mutation confers a dominant negative via the Quickchange mutagenesis kit (Stratagene). The mutation phenotype. Insert size = 660 bp. confers a dominant negative phenotype. Insert size = 560 bp. RASK20TND0 K-Ras2 S17N 3xHA-tagged(N-terminus) RAP1B000D0 RAP1B S17N N-terminal 3x-hemagglutinin tagged human Ras family small Human Ras family small GTP binding protein RAP1B S17N GTP binding protein K-Ras2 (S17N mutant) cloned into (dominant negative) cloned into pcDNA3.1+ (Invitrogen) at pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N BamHI (5') and XhoI (3'). The S17N mutation was introduced into mutation was introduced into K-Ras2 3X HA tagged (N- the Ras family small GTP binding protein Rap1B (RAP1B00000) terminus)(RASK20TN00) via the Quickchange mutagenesis kit via the Quickchange mutagenesis kit (Stratagene). The mutation (Stratagene). The mutation confers a dominant negative confers a dominant negative phenotype. Insert size = 560 bp. phenotype. Insert size = 660 bp.

RASH0000D0 H-Ras S17N RASM00TND0 M-Ras S27N 3xHA-tagged(N-terminus) The S17N mutation was introduced into the Ras family small N-terminal 3x-hemagglutinin tagged human Ras family small GTP binding protein H-Ras (RASH000000) via the Quickchange GTP binding protein M-Ras (S27N mutant) cloned into mutagenesis kit (Stratagene). The mutation confers a dominant pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S27N negative phenotype. Insert size = 570 bp. mutation was introduced into M-Ras 3X HA tagged (N- terminus)(RASM00TN00) via the Quickchange mutagenesis kit RASK2000D0 K-Ras2 S17N (Stratagene). The mutation confers a dominant negative The S17N mutation was introduced into the Ras family small phenotype. Insert size = 720 bp. GTP binding protein K-Ras2 (RASK200000) via the Quickchange mutagenesis kit (Stratagene). The mutation RASN00TND0 N-Ras S17N 3xHA-tagged(N-terminus) confers a dominant negative phenotype. Insert size = 570 bp. N-terminal 3x-hemagglutinin tagged human Ras family small GTP binding protein N-Ras (S17N mutant) cloned into RASM0000D0 M-Ras S27N pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N The S27N mutation was introduced into the Ras family small mutation was introduced into N-Ras 3X HA tagged (N- GTP binding protein M-Ras (RASM000000) via the Quickchange terminus)(RASN00TN00) via the Quickchange mutagenesis kit mutagenesis kit (Stratagene). The mutation confers a dominant (Stratagene). The mutation confers a dominant negative negative phenotype. Insert size = 630 bp. phenotype. Insert size = 670 bp.

RASN0000D0 N-Ras S17N RASR00TND0 R-Ras S43N 3xHA-tagged(N-terminus) The S17N mutation was introduced into the Ras family small N-terminal 3x-hemagglutinin tagged human Ras family small GTP binding protein N-Ras (RASN000000) via the Quickchange GTP binding protein R-Ras (S43N mutant) cloned into mutagenesis kit (Stratagene). The mutation confers a dominant pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S43N negative phenotype. Insert size = 580 bp. mutation was introduced into R-Ras 3X HA tagged (N- terminus)(RASR00TN00) via the Quickchange mutagenesis kit RASR0000D0 R-Ras S43N (Stratagene). The mutation confers a dominant negative The S43N mutation was introduced into the Ras family small phenotype. Insert size = 750 bp. GTP binding protein R-Ras (RASR000000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a dominant negative phenotype. Insert size = 660 bp. Ras Family Wild-Type 3xHA Tagged

RAP1A0TN00 RAP1A 3XHA-tagged (N-terminus) Ras Family SN MUTANTS 3xHA Tagged N-terminal 3XHemagglutinin-tagged human Ras family small GTP binding protein RAP1A (wild type) cloned into pcDNA3.1+ RAP1A0TND0 RAP1A S17N 3XHA-tagged (N-terminus) (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame N-terminal 3XHemagglutinin-tagged human Ras family small was amplified by the PCR from human whole brain cDNA GTP binding protein RAP1A S17N (dominant negative) cloned (Clontech). The insert was sequenced and found to be identical into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N with GB ACC#X12533. Insert size = 640 bp. mutation was introduced into the Ras family small GTP binding protein Rap1A (RAP1A00000) via the Quickchange mutagenesis RAP1B0TN00 RAP1B 3XHA-tagged (N-terminus) kit (Stratagene). The mutation confers a dominant negative N-terminal 3Xhemagglutinin-tagged human Ras family small phenotype. Insert size = 640 bp. GTP binding protein RAP1B (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame RAP1B0TND0 RAP1B S17N 3XHA-tagged (N-terminus) was amplified by the PCR from human whole brain cDNA N-terminal 3XHemagglutinin-tagged human Ras family small (Clontech). The insert was sequenced and found to be identical with GB ACC#X08004. Insert size = 640 bp.

48 cDNA Resource Center

RASH00TN00 H-Ras 3xHA-tagged(N-terminus) 2XMYC SN N-terminal 3X-hemagglutinin tagged human Ras family small GTP binding protein H-Ras (wild type) cloned into pcDNA3.1+ RAP1A0MND0 RAP1A S17N 2XMYC-tagged (N-terminus) (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame N-terminal 2XMYC-tagged human Ras family small GTP binding was derived from RASH000000 by the PCR. Insert = 670 bp. protein RAP1A S17N (dominant negative) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N RASK20TN00 K-Ras2 3xHA-tagged(N-terminus) mutation was introduced into the Ras family small GTP binding N-terminal 3X-hemagglutinin tagged human Ras family small protein Rap1A (RAP1A00000) via the Quickchange mutagenesis GTP binding protein K-Ras2 (wild type) cloned into pcDNA3.1+ kit (Stratagene). The mutation confers a dominant negative (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame phenotype. Insert size = 640 bp. was derived from RASK200000 by the PCR. Insert size = 670 bp. RAP1B0MND0 RAP1B S17N 2XMYC-tagged (N-terminus) RASM00TN00 M-Ras 3xHA-tagged(N-terminus) N-terminal 2XMYC-tagged human Ras family small GTP binding N-terminal 3X-hemagglutinin tagged human Ras family small protein RAP1B S17N (dominant negative) cloned into GTP binding protein M-Ras (wild type) cloned into pcDNA3.1+ pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame mutation was introduced into the Ras family small GTP binding was derived from RASM000000 by the PCR. Insert size = 730 protein Rap1B (RAP1B00000) via the Quickchange mutagenesis bp. kit (Stratagene). The mutation confers a dominant negative phenotype. Insert size = 640 bp. RASN00TN00 N-Ras 3xHA-tagged(N-terminus) N-terminal 3X-hemagglutinin tagged human Ras family small RASH00MND0 H-Ras S17N 2XMYC-tagged(N-terminus) GTP binding protein N-Ras (wild type) cloned into pcDNA3.1+ N-terminal 2XMYC-tagged human Ras family small GTP binding (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame protein H-Ras S17N (dominant negative) cloned into pcDNA3.1+ was derived from RASN000000 by the PCR. Insert size = 680 (Invitrogen) at KpnI (5') and XhoI (3'). The S17N mutation was bp. introduced into the Ras family small GTP binding protein H-Ras (RASH000000) via the Quickchange mutagenesis kit RASR00TN00 R-Ras 3xHA-tagged(N-terminus) (Stratagene). The mutation confers a dominant negative N-terminal 3X-hemagglutinin tagged human Ras family small phenotype. Insert size = 670 bp. GTP binding protein R-Ras (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame RASK20MND0 K-Ras2 S17N 2XMYC-tagged(N-terminus) was derived from RASR000000 by the PCR. Insert size = 760 N-terminal 2XMYC- tagged human Ras family small GTP binding bp. protein K-Ras2 S17N (dominant negative) cloned into

pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The S17N 2XMYC GV mutation was introduced into the Ras family small GTP binding protein K-Ras2 (RASK200000) via the Quickchange RAP1A0MNC0 RAP1A G12V 2XMYC-tagged (N-terminus) mutagenesis kit (Stratagene). The mutation confers a dominant N-terminal 2XMYC-tagged human Ras family small GTP binding negative phenotype. . Insert size = 670 bp. protein RAP1A G12V (constitutively active) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V 2XMYC-tagged mutation was introduced into the Ras family small GTP binding protein Rap1A (RAP1A00000) via the Quickchange mutagenesis RAP1A0MN00 RAP1A 2XMYC-tagged (N-terminus) kit (Stratagene). The mutation confers a constitutively active N-terminal 2XMYC-tagged human Ras family small GTP binding phenotype. Insert size = 640 bp. protein RAP1A (wild type) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was amplified by RAP1B0MNC0 RAP1B G12V 2XMYC-tagged (N-terminus) the PCR from human whole brain cDNA (Clontech). The insert N-terminal 2XMYC-tagged human Ras family small GTP binding was sequenced and found to be identical with GB ACC#X12533. protein RAP1B G12V (constitutively active) cloned into Insert size = 640 bp. pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V mutation was introduced into the Ras family small GTP binding RAP1B0MN00 RAP1B 2XMYC-tagged (N-terminus) protein Rap1B (RAP1B00000) via the Quickchange mutagenesis N-terminal 2XMYC-tagged human Ras family small GTP binding kit (Stratagene). The mutation confers a constitutively active protein RAP1B (wild type) cloned into pcDNA3.1+ (Invitrogen) at phenotype. Insert size = 640 bp. KpnI (5') and XhoI (3'). The open reading frame was amplified by the PCR from human whole brain cDNA (Clontech). The insert RASH00MNC0 H-Ras G12V 2XMYC-tagged(N-terminus) was sequenced and found to be identical with GB ACC#X08004. N-terminal 2XMYC-tagged human Ras family small GTP binding Insert size = 640 bp. protein H-Ras G12V (constitutively active) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V RASH00MN00 H-Ras 2XMYC-tagged(N-terminus) mutation was introduced into the Ras family small GTP binding N-terminal 2XMYC-tagged human Ras family small GTP binding protein H-Ras (RASH000000) via the Quickchange mutagenesis protein H-Ras (wild type) cloned into pcDNA3.1+ (Invitrogen) at kit (Stratagene). The mutation confers a constitutively active KpnI (5') and XhoI (3'). The open reading frame was derived phenotype. Insert size = 670 bp. from RASH000000 by the PCR. Insert size = 670 bp.

RASK20MNC0 K-Ras2 G12V 2XMYC-tagged(N-terminus) RASK20MN00 K-Ras2 2XMYC-tagged (N-terminus) N-terminal 2XMYC- tagged human Ras family small GTP binding N-terminal 2XMYC- tagged human Ras family small GTP binding protein K-Ras2 G12V (constitutively active) cloned into protein K-Ras2 (wild type) cloned into pcDNA3.1+ (Invitrogen) at pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V KpnI (5') and XhoI (3'). The open reading frame was derived mutation was introduced into the Ras family small GTP binding from RASK200000 by the PCR. Insert size = 670 bp. protein K-Ras2 (RASK200000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active phenotype. . Insert size = 670 bp.

49 cDNA Resource Center

Ras-like G-proteins

Wild-Type Native Ras-like proteins Wild-Type in DNR-1r Native Ras-like proteins in pDNR-1r vector

Ras-like Wild-Type TC21000000 TC21 GenBank Accession # AF493924: Human Ras family small AGS0100000 AGS1 GTP binding protein TC21 (wild type) cloned into pcDNA3.1+ GenBank Accession # AF498923: Human activator of G-protein (Invitrogen) at BamH I (5') and Xho I (3'). The open reading signaling AGS1 (wild type) cloned into pcDNA3.1+ (Invitrogen) frame was amplified by the PCR from human heart cDNA at BamHI (5') and XhoI (3'). The open reading frame was (Clontech). The insert was sequenced. Insert size = 620 bp. amplified by the PCR from human whole brain cDNA (Clontech). The insert was sequenced. Insert size = 850 bp. Ras-like Wild-Type in DNR-1r RALA000000 RALA GenBank Accession # AF493910: Human Ras family small GS0100001 AGS1 (pDNR-1r) GTP binding protein RALA (wild type) cloned into pcDNA3.1+ Human activator of G-protein signaling AGS1 (wild type) (Invitrogen) at BamH I (5') and Xho I (3'). The open reading AGS0100000 cloned into pDNR-1r (Clontech) at SmaI (5') and frame was amplified by the PCR from human lung cDNA XhoI (3'). The SmaI site was destroyed(* ). The insert can be (Clontech). The insert was sequenced. Insert size = 635 bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical RALB000000 RALB with GB ACC # AF498923. Insert size = 850 bp. GenBank Accession # AF493911: Human Ras family small GTP binding protein RALB (wild type) cloned into pcDNA3.1+ RALA000001 RALA (pDNR-1r) (Invitrogen) at BamH I (5') and Xho I (3'). The open reading Human Ras family small GTP binding protein RALA (wild type) frame was amplified by the PCR from human whole brain RALA000000 cloned into pDNR-1r (Clontech) at SmaI (5') and cDNA (Clontech). The insert was sequenced. Insert size = 625 XhoI (3'). The SmaI site was destroyed(*). The insert can be bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical RHEB200000 RHEB2 with GB ACC # AF493910. Insert size = 635 bp. GenBank Accession # AF493921: Human Ras family small GTP binding protein RHEB2 (wild type) cloned into pcDNA3.1+ RHEB200001 RHEB2 (pDNR-1r) (Invitrogen) at BamH I (5') and Xho I (3'). The open reading Human Ras family small GTP binding protein RHEB2 (wild type) frame was amplified by the PCR from human whole brain RHEB200000 cloned into pDNR-1r (Clontech) at SmaI (5') and cDNA (Clontech). The insert was sequenced. Insert size = 565 XhoI (3'). The SmaI site was destroyed(*). The insert can be bp. excised with SalI and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical RIG0000000 Rig with GB ACC # AF493921. Insert size = 565 bp. GeneBank Accession # AY180973. Human Ras family small GTP binding protein Rig (wild type) cloned into pcDNA3.1+ RIN0000001 RIN (pDNR-1r) (Invitrogen) at EcoRI (5') and XhoI (3'). The open reading frame Human Ras family small GTP binding protein RIN (wild type) was amplified by the PCR from human whole brain cDNA RIN0000000 cloned into pDNR-1r (Clontech) at SmaI (5') and (Clontech) . The insert was sequenced. Insert size= 597 XhoI (3'). The SmaI site was destroyed(*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be RIN0000000 RIN present. The insert was sequenced and found to be identical GenBank Accession # AF493922: Human Ras family small with GB ACC # AF493922. Insert size = 665 bp. GTP binding protein RIN (wild type) cloned into pcDNA3.1+ (Invitrogen) at BamHI (5') and XhoI (3'). The open reading RIT0000001 RIT (pDNR-1r) frame was amplified by the PCR from IMAGE clone (45534). Human Ras family small GTP binding protein RIT (wild type) The insert was sequenced. Insert size = 665 bp. RIT0000000 cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(*). The insert can be RIT0000000 RIT excised with SalI and the 3’ RE. Internal SalI sites may be GenBank Accession # AF493923: Human Ras family small present. The insert was sequenced and found to be identical GTP binding protein RIT (wild type) cloned into pcDNA3.1+ with GB ACC # AF493923. Insert size = 670 bp. (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (268332). The insert was sequenced. Insert size = 670 bp.

50 cDNA Resource Center

Small G-proteins Rho family

Wild-Type Native Rho family proteins pDNR-1r Native Rho family proteins in pDNR-1r vector 3xHA-Tagged Native Rho family proteins with an N-terminal hemagglutinin tag in triplicate GV Mutants Constitutively active phenotype 3xHA GV Rho family proteins with a constitutively active phenotype and an N-terminal hemagglutinin tag in triplicate TN Mutants Rho family proteins with a dominant negative phenotype due to a threonine (T) to asparagine (N) mutation 3xHA TN Rho family proteins with a constitutively active phenotype and an N-terminal hemagglutinin tag in triplicate 2XMYC GV Rho family proteins with a constitutively active phenotype and an N-terminal MYC tag in duplicate 2XMYC TN Rho family proteins with a dominant negative phenotype and an N-terminal MYC tag in duplicate 2XMYC-tagged Native Rho family proteins with an N-terminal MYC tag in duplicate GV PBRQ 2XMYC Rho family proteins with a constitutively active phenotype, the polybasic region mutated, and an N-terminal MYC tag in duplicate GV PBRQ 3XHA Rho family proteins with a constitutively active phenotype, the polybasic region mutated, and an N-terminal hemagglutinin tag in triplicate PBRQ 2XMYC Rho family proteins with the polybasic region mutated and an N-terminal MYC tag in duplicate PBRQ 3XHA Rho family proteins with the polybasic region mutated and an N-terminal hemagglutinin tag in triplicate TN PBRQ 2XMYC Rho family proteins with a dominant negative phenotype, the polybasic region mutated, and an N-terminal MYC tag in duplicate TN PBRQ 3XHA Rho family proteins with a dominant negative phenotype, the polybasic region mutated, and an N-terminal hemagglutinin tag in triplicate

(Clontech). The insert was sequenced. Insert size = 700 bp. Rho Family Wild-Type RHO0700000 Rho7 (Rnd2) CDC4200000 CDC42 GenBank Accession # AF498968: Human Rho family small GTP GenBank Accession # AF498962: Human Rho family small GTP binding protein Rho7 (Rnd2) (wild type) cloned into pcDNA3.1+ binding protein CDC42 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame (Invitrogen) at EcoRI (5') and Xba I (3'). The open reading frame was amplified by the PCR from IMAGE clone (1650500). The was amplified by the PCR from human whole brain cDNA insert was sequenced. Insert size = 690 bp. (Clontech). The insert was sequenced. Insert size = 600 bp. RHO0800000 Rho8 (RhoE, Rnd3) CDC42P0000 CDC42 placental isoform GenBank Accession # AF498969: Human Rho family small GTP GenBank Accession # AF498963: Human Rho family small GTP binding protein Rho8 (Rnd3, RhoE) (wild type) cloned into binding protein CDC42 placental isoform (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame was amplified by the PCR from human placental reading frame was amplified by the PCR from human placental cDNA (Clontech). The insert was sequenced. Insert size = 745 cDNA (Clontech). The insert was sequenced. Insert size = 600 bp. bp. RHO0A00000 RhoA RAC0100000 Rac1 GenBank Accession # AF498970: Human Rho family small GTP GenBank Accession # AF498964: Human Rho family small GTP binding protein RhoA (wild type) cloned into pcDNA3.1+ binding protein Rac1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from IMAGE clone (1657288). The (Clontech). The insert was sequenced. Insert size = 600 bp. insert was sequenced. Insert size = 630 bp. RHO0B00000 RhoB RAC0200000 Rac2 GenBank Accession # AF498971: Human Rho family small GTP GenBank Accession # AF498965: Human Rho family small GTP binding protein RhoB (wild type) cloned into pcDNA3.1+ binding protein Rac2 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xba I (3'). The open reading frame (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from IMAGE clone (245761). The (Clontech). The insert was sequenced. Insert size = 600 bp. insert was sequenced. Insert size = 575bp. RHO0C00000 RhoC RAC0300000 Rac3 GenBank Accession # AF498972: Human Rho family small GTP GenBank Accession # AF498966: Human Rho family small GTP binding protein RhoC (wild type) cloned into pcDNA3.1+ binding protein Rac3 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame was amplified by the PCR from human whole brain cDNA was amplified by the PCR from IMAGE clone (1739912). The (Clontech). The insert was sequenced. Insert size = 600 bp. insert was sequenced. Insert size = 590 bp. RHO0D00000 RhoD (RhoHP1) RHO0600000 Rho6 GenBank Accession # AF498973: Human Rho family small GTP GenBank Accession # AF498967: Human Rho family small GTP binding protein RhoD (wild type) cloned into pcDNA3.1+ binding protein Rho6 (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xba I (3'). The open reading frame (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame was amplified by the PCR from IMAGE clone (1565600). The was amplified by the PCR from human whole brain cDNA insert was sequenced. Insert size = 640 bp.

51 cDNA Resource Center

RHO0G00000 RhoG RHO0600001 Rho6 (pDNR-1r) GenBank Accession # AF498974: Human Rho family small GTP Human Rho family small GTP binding protein Rho6 (wild type) binding protein RhoG (wild type) cloned into pcDNA3.1+ cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame SmaI site was destroyed (* ). The insert can be excised with SalI was amplified by the PCR from human whole brain cDNA and the 3’ RE. Internal SalI sites may be present. The insert was (Clontech). The insert was sequenced. Insert size = 580 bp. sequenced and found to be identical with GB ACC # AF498967. Insert size = 700 bp. RHO0H00000 RhoH (TTF) GenBank Accession # AF498975: Human Rho family small GTP RHO0700001 Rho7 (Rnd2) (pDNR-1r) binding protein RhoH (TTF) (wild type) cloned into pcDNA3.1+ Human Rho family small GTP binding protein Rho7 (Rnd2) (wild (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame type) cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). was amplified by the PCR from IMAGE clone (2021711). The The SmaI site was destroyed (* ). The insert can be excised with insert was sequenced. Insert size = 580 bp. SaII and the 3’ RE. Internal SalI sites may be present. The insert was sequenced and found to be identical with GB ACC # TC10000000 TC10 AF498968. Insert size = 690 bp. GenBank Accession # AF498976: Human Rho family small GTP binding protein TC10 (wild type) cloned into pcDNA3.1+ RHO0800001 Rho8 (RhoE, Rnd3) (pDNR-1r) (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame Human Rho family small GTP binding protein Rho8 (Rnd3, was amplified by the PCR from human whole brain cDNA RhoE) (wild type) cloned into pDNR-1r (Clontech) at SmaI (5') (Clontech). The insert was sequenced. Insert size = 650 bp. and XhoI (3'). The SmaI site was destroyed (*). The insert can be excised with SalI and the 3’ RE. Internal SalI sites may be TCL0000000 TCL present. The insert was sequenced and found to be identical with GenBank Accession # AF498977: Human Rho family small GTP GB ACC # AF498969. Insert size = 745 bp. binding protein TCL (wild type) cloned into pcDNA3.1+ (Invitrogen) at EcoRI (5') and Xho I (3'). The open reading frame RHO0A00001 RhoA (pDNR-1r) was amplified by the PCR from IMAGE clone (1682327). The Human Rho family small GTP binding protein RhoA (wild type) insert was sequenced. Insert size = 660 bp. cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The SmaI site was destroyed(* ). The insert can be excised with SalI and the 3” RE. Internal SalI sites may be present. The insert was Rho Family Wild-Type in pDNR-1r sequenced and found to be identical with GB ACC# AF498970. Insert size = 600 bp. CDC4200001 CDC42 (pDNR-1r) Human Rho family small GTP binding protein CDC42 (wild type) RHO0B00001 RhoB (pDNR-1r) cloned into pDNR-1r (Clontech) at SmaI (5') and XbaI (3'). The Human Rho family small GTP binding protein RhoB (wild type) SmaI site was destroyed (* ). The insert can be excised with SalI cloned into pDNR-1r (Clontech) at SmaI (5') and XbaI (3'). The and the 3’ RE. Internal SalI sites may be present. The insert was SmaI site was destroyed(* ). The insert can beexcised with SalI sequenced and found to be identical with GB ACC # AF498962. and the 3' RE. Internal SalI sites may be present. The insert was Insert size = 600 bp. sequenced and found to be identical with GB ACC # AF498971. Insert size = 600 bp. CDC42P0001 CDC42 placental isoform (pDNR-1r) Human Rho family small GTP binding protein CDC42 placental RHO0C00001 RhoC (pDNR-1r) isoform (wild type) cloned into pDNR-1r (Clontech) at SmaI (5') Human Rho family small GTP binding protein RhoC (wild type) and XhoI (3'). The SmaI site was destroyed (*). The insert can be cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The excised with SalI and the 3’ RE. Internal SalI sites may be SmaI site was destroyed(* ). The insert can beexcised with SalI present. The insert was sequenced and found to be identical with and the 3’ RE. Internal SalI sites may be present. The insert was GB ACC # AF498963. Insert size = 600 bp. sequenced and found to be identical with GB ACC # AF498972. Insert size = 600 bp. RAC0100001 Rac1 (pDNR-1r) Human Rho family small GTP binding protein Rac1 (wild type) RHO0D00001 RhoD (RhoHP1) (pDNR-1r) cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The Human Rho family small GTP binding protein RhoD (HP1) (wild SmaI site was destroyed (* ). The insert can be excised with SalI type) cloned into pDNR-1r (Clontech) at SmaI (5') and XbaI (3'). and the 3’ RE. Internal SalI sites may be present. The insert was The SmaI site was destroyed(* ). The insert can beexcised with sequenced and found to be identical with GB ACC # AF498964. SalI and the 3’ RE. Internal SalI sites may be present. The insert Insert size = 630 bp. was sequenced and found to be identical with GB ACC # AF498973. Insert size = 640 bp. RAC0200001 Rac2 (pDNR-1r) Human Rho family small GTP binding protein Rac2 (wild type) RHO0G00001 RhoG (pDNR-1r) cloned into pDNR-1R (Clontech) at SmaI (5') and XhoI (3'). The Human Rho family small GTP binding protein RhoG (wild type) SmaI site was destroyed (* ). The insert can be excised with SalI cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The and the 3’ RE. Internal SalI sites may be present. The insert was SmaI site was destroyed(* ). The insert can be excised with SalI sequenced and found to be identical with GB ACC # AF498965. and the 3’ RE. Internal SalI sites may be present. The insert was Insert size = 575 bp. sequenced and found to be identical with GB ACC # AF498974. Insert size = 580 bp. RAC0300001 Rac3 (pDNR-1r) Human Rho family small GTP binding protein Rac3 (wild type) RHO0H00001 RhoH (TTF) (pDNR-1r) cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The Human Rho family small GTP binding protein RhoH (TTF) (wild SmaI site was destroyed(* ). The insert can be excised with SalI type) cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). and the 3’RE. The insert was sequenced and found to be The SmaI site was destroyed(* ). The insert can be excised with identical with GB ACC # AF498966. Insert size = 590 bp. SalI and the 3’ RE. Internal SalI sites may be present. The insert

52 cDNA Resource Center

was sequenced and found to be identical with GB ACC # was derived from RHO0B00000 by the PCR. Insert size = 680 AF498975. Insert size = 580 bp. bp.

TC10000001 TC10 (pDNR-1r) RHO0C0TN00 RhoC 3XHA-tagged(N-terminus) Human Rho family small GTP binding protein TC10 (wild type) N-terminal 3X-hemagglutinin tagged human Rho family small cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The GTP binding protein RhoC (wild type) cloned into pcDNA3.1+ SmaI site was destroyed (* ). The insert can be excised with SalI (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame and the 3’RE. Internal SalI sites may be present. The insert was was derived from RHO0C00000 by the PCR. Insert size = 680 sequenced and found to be identical with GB ACC # AF498976. bp. Insert size = 650 bp.

TCL0000001 TCL (pDNR-1r) Rho GV Mutants Human Rho family small GTP binding protein TCL (wild type) cloned into pDNR-1r (Clontech) at SmaI (5') and XhoI (3'). The CDC42000C0 CDC42 G12V SmaI site was destroyed (* ). The insert can be excised with SalI The G12V mutation was introduced into the Rho family small and te3’ RE. Internal SalI sites may be present. The insert was GTP binding protein CDC42 (CDC4200000) via the Quickchange sequenced and found to be identical with GB ACC # AF498977. mutagenesis kit (Stratagene). The mutation confers a Insert size = 660 bp. constitutively active phenotype. Insert size = 600 bp.

CDC42P00C0 CDC42 placental isoform G12V Rho Family Wild-Type 3xHA-Tagged The G12V mutation was introduced into the Rho family small GTP binding protein CDC42 placental isoform (CDC42P0000) CDC420TN00 CDC42 3XHA-tagged(N-terminal) via the Quickchange mutagenesis kit (Stratagene). The mutation N-terminal 3X-hemagglutinin tagged human Rho family small confers a constitutively active phenotype. Insert size = 600 bp. GTP binding protein CDC42 (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I (5') and Xba I (3'). The open reading frame RAC01000C0 Rac1 G12V was derived from CDC4200000 by the PCR. Insert size = 670 The G12V mutation was introduced into the Rho family small bp. GTP binding protein Rac1 (RAC0100000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a CDC42PTN00 CDC42 placental isoform 3XHA-tagged(N- constitutively active phenotype. Insert size = 630 bp. terminus) N-terminal 3X-hemagglutinin tagged human Rho family small RAC02000C0 Rac2 G12V GTP binding protein CDC42 placental isoform (wild type) cloned The G12V mutation was introduced into the Rho family small into pcDNA3.1+ (Invitrogen) at Kpn I (5') and Xho I (3'). The GTP binding protein Rac2 (RAC0200000) via the Quickchange open reading frame was derived from CDC42P0000 by the PCR. mutagenesis kit (Stratagene). The mutation confers a Insert size = 670 bp. constitutively active phenotype. Insert size = 575 bp.

RAC010TN00 Rac1 3XHA-tagged(N-terminus) RAC03000C0 Rac3 G12V N-terminal 3X-hemagglutinin tagged human Rho family small The G12V mutation was introduced into the Rho family small GTP binding protein Rac1 (wild type) cloned into pcDNA3.1+ GTP binding protein Rac3 (RAC0300000) via the Quickchange (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame mutagenesis kit (Stratagene). The mutation confers a was derived from RAC0100000 by the PCR. Insert size = 730 constitutively active phenotype. Insert size = 590 bp. bp. RHO0A000C0 RhoA G14V RAC020TN00 Rac2 3XHA-tagged(N-terminus) The G14V mutation was introduced into the Rho family small N-terminal 3X-hemagglutinin tagged human Rho family small GTP binding protein RhoA (RHO0A00000) via the Quickchange GTP binding protein Rac2 (wild type) cloned into pcDNA3.1+ mutagenesis kit (Stratagene). The mutation confers a (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame constitutively active phenotype. Insert size = 600 bp. was derived from RAC0200000 by the PCR. Insert size = 680 bp. RHO0B000C0 RhoB G14V The G14V mutation was introduced into the Rho family small RAC030TN00 Rac3 3xHA-tagged(N-terminus) GTP binding protein RhoB (RHO0B00000) via the Quickchange N-terminal 3X-hemagglutinin tagged human Rho family small mutagenesis kit (Stratagene). The mutation confers a GTP binding protein Rac3 (wild type) cloned into pcDNA3.1+ constitutively active phenotype. Insert size = 600 bp. (Invitrogen) at KpnI (5') and XhoI (3'). The open reading frame was derived from RAC0300000 by the PCR. Insert size = 680 RHO0C000C0 RhoC G14V bp. The G14V mutation was introduced into the Rho family small GTP binding protein RhoC (RHO0C00000) via the Quickchange RHO0A0TN00 RhoA 3XHA-tagged(N-terminus) mutagenesis kit (Stratagene). The mutation confers a N-terminal 3X-hemagglutinin tagged human Rho family small constitutively active phenotype. Insert size = 600 bp. GTP binding protein RhoA (wild type) cloned into pcDNA3.1+ (Invitrogen) at Kpn I (5') and Xho I (3'). The open reading frame was derived from RHO0A00000 by the PCR. Insert size = 680 Rho GV Mutants 3xHA-Tagged bp. CDC420TNC0 CDC42 G12V 3xHA-tagged (N-terminal) RHO0B0TN00 RhoB 3XHA-tagged(N-terminus) N-terminal 3x-hemagglutinin tagged human Rho family small N-terminal 3X-hemagglutinin tagged human Rho family small GTP binding protein CDC42 (G12V mutant) cloned into GTP binding protein RhoB (wild type) cloned into pcDNA3.1+ pcDNA3.1+ (Invitrogen) at KpnI (5') and XbaI (3'). The G12V (Invitrogen) at Kpn I (5') and Xba I (3'). The open reading frame mutation was introduced into CDC42 3X HA tagged (N-

53 cDNA Resource Center terminus)(CDC420TN00) via the Quickchange mutagenesis kit Rho TN Mutants (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 670 bp. CDC42000D0 CDC42 T17N The T17N mutation was introduced into the Rho family small CDC42PTNC0 CDC42 placental isoform G12V 3xHA-tagged GTP binding protein CDC42 (CDC4200000) via the Quickchange (N-terminus) mutagenesis kit (Stratagene). The mutation confers a dominant N-terminal 3x-hemagglutinin tagged human Rho family small negative phenotype. Insert size = 600 bp. GTP binding protein CDC42 placental isoform (G12V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). CDC42P00D0 CDC42 placental isoform T17N The G12V mutation was introduced into CDC42 placental The T17N mutation was introduced into the Rho family small isoform 3X HA tagged (N-terminus)(CDC42PTN00) via the GTP binding protein CDC42 placental isoform (CDC42P0000) Quickchange mutagenesis kit (Stratagene). The mutation via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active phenotype. Insert size = 670 bp. confers a dominant negative phenotype. Insert size = 600 bp.

RAC010TNC0 Rac1 G12V 3xHA-tagged(N-terminus) RAC01000D0 Rac1 T17N N-terminal 3x-hemagglutinin tagged human Rho family small The T17N mutation was introduced into the Rho family small GTP binding protein Rac1 (G12V mutant) cloned into GTP binding protein Rac1 (RAC0100000) via the Quickchange pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V mutagenesis kit (Stratagene). The mutation confers a dominant mutation was introduced into Rac1 3X HA tagged (N- negative phenotype. Insert size = 630 bp. terminus)(RAC010TN00) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active RAC02000D0 Rac2 T17N phenotype. Insert size = 730 bp. The T17N mutation was introduced into the Rho family small GTP binding protein Rac2 (RAC0200000) via the Quickchange RAC020TNC0 Rac2 G12V 3xHA-tagged(N-terminus) mutagenesis kit (Stratagene). The mutation confers a dominant N-terminal 3x-hemagglutinin tagged human Rho family small negative phenotype. Insert size = 575 bp. GTP binding protein Rac2 (G12V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V RAC03000D0 Rac3 T17N mutation was introduced into Rac2 3X HA tagged (N- The T17N mutation was introduced into the Rho family small terminus)(RAC020TN00) via the Quickchange mutagenesis kit GTP binding protein Rac3 (RAC0300000) via the Quickchange (Stratagene). The mutation confers a constitutively active mutagenesis kit (Stratagene). The mutation confers a dominant phenotype. Insert size = 680 bp. negative phenotype. Insert size = 590 bp.

RAC030TNC0 Rac3 G12V 3xHA-tagged(N-terminus) RHO0A000D0 RhoA T19N N-terminal 3x-hemagglutinin tagged human Rho family small The T19N mutation was introduced into the Rho family small GTP binding protein Rac3 (G12V mutant) cloned into GTP binding protein RhoA (RHO0A00000) via the Quickchange pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G12V mutagenesis kit (Stratagene). The mutation confers a dominant mutation was introduced into Rac3 3X HA tagged (N- negative phenotype. Insert size = 600 bp. terminus)(RAC030TN00) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active RHO0B000D0 RhoB T19N phenotype. Insert size = 670 bp. The T19N mutation was introduced into the Rho family small GTP binding protein RhoB (RHO0B00000) via the Quickchange RHO0A0TNC0 RhoA G14V 3xHA-tagged(N-terminus) mutagenesis kit (Stratagene). The mutation confers a dominant N-terminal 3x-hemagglutinin tagged human Rho family small negative phenotype. Insert size = 600 bp. GTP binding protein RhoA (G14V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G14V RHO0C000D0 RhoC T19N mutation was introduced into RhoA 3X HA tagged (N- The T19N mutation was introduced into the Rho family small terminus)(RHO0A0TN00) via the Quickchange mutagenesis kit GTP binding protein RhoC (RHO0C00000) via the Quickchange (Stratagene). The mutation confers a constitutively active mutagenesis kit (Stratagene). The mutation confers a dominant phenotype. Insert size = 680 bp. negative phenotype. Insert size = 600 bp.

RHO0B0TNC0 RhoB G14V 3xHA-tagged(N-terminus) N-terminal 3x-hemagglutinin tagged human Rho family small Rho TN Mutants 3xHA-Tagged GTP binding protein RhoB (G14V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XbaI (3'). The G14V CDC420TND0 CDC42 T17N 3xHA-tagged(N-terminus) mutation was introduced into RhoB 3X HA tagged (N- N-terminal 3x-hemagglutinin tagged human Rho family small terminus)(RHO0B0TN00) via the Quickchange mutagenesis kit GTP binding protein CDC42 (T17N mutant) cloned into (Stratagene). The mutation confers a constitutively active pcDNA3.1+ (Invitrogen) at KpnI (5') and XbaI (3'). The T17N phenotype. Insert size = 680 bp. mutation was introduced into CDC42 3X HA tagged (N-

terminus)(CDC420TN00) via the Quickchange mutagenesis kit RHO0C0TNC0 RhoC G14V 3xHA-tagged(N-terminus) (Stratagene). The mutation confers a dominant negative N-terminal 3x-hemagglutinin tagged human Rho family small phenotype. Insert size = 690 bp. GTP binding protein RhoC (G14V mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The G14V CDC42PTND0 CDC42 placental isoform T17N 3xHA- mutation was introduced into RhoC 3X HA tagged (N- tagged(N-terminus) terminus)(RHO0C0TN00) via the Quickchange mutagenesis kit N-terminal 3x-hemagglutinin tagged human Rho family small (Stratagene). The mutation confers a constitutively active GTP binding protein CDC42 placental isoform (T17N mutant) phenotype. Insert size = 680 bp. cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3').

The T17N mutation was introduced into CDC42 placental

isoform 3X HA tagged (N-terminus)(CDC42PTN00) via the

54 cDNA Resource Center

Quickchange mutagenesis kit (Stratagene). The mutation introduced into the Rho family small GTP binding protein RhoA confers a dominant negative phenotype. Insert size = 690 bp. (RHO0A00000) via the Quickchange mutagenesis kit (Stratagene). The mutation confers a constitutively active RAC010TND0 Rac1 T17N 3xHA-tagged(N-terminus) phenotype. Insert size = 680 bp. N-terminal 3x-hemagglutinin tagged human Rho family small GTP binding protein Rac1 (T17N mutant) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The T17N mutation was 2XMYC TN introduced into Rac1 3X HA tagged (N-terminus)(RAC010TN00) via the Quickchange mutagenesis kit (Stratagene). The mutation RAC010MND0 Rac1 T17N 2XMYC-tagged (N-terminus) confers a dominant negative phenotype. Insert size = 720 bp. N-terminal 2XMYC-tagged human Rho family small GTP binding protein Rac1 T17N (dominant negative) cloned into pcDNA3.1+ RAC020TND0 Rac2 T17N 3xHA-tagged(N-terminus) (Invitrogen) at KpnI (5') and XhoI (3'). The T17N mutation was N-terminal 3x-hemagglutinin tagged human Rho family small introduced into the Rho family small GTP binding protein Rac1 GTP binding protein Rac2 (T17N mutant) cloned into pcDNA3.1+ (RAC0100000) via the Quickchange mutagenesis kit (Invitrogen) at KpnI (5') and XhoI (3'). The T17N mutation was (Stratagene). The mutation confers a dominant negative introduced into Rac2 3X HA tagged (N-terminus)(RAC020TN00) phenotype. Insert size = 720 bp. via the Quickchange mutagenesis kit (Stratagene). The mutation confers a dominant negative phenotype. Insert size = 670 bp. RHO0A0MND0 RhoA T19N 2XMYC-tagged (N-terminus) N-terminal 2XMYC-tagged human Rho family small GTP binding RAC030TND0 Rac3 T17N 3xHA-tagged(N-terminus) protein RhoA T19N (dominant negative) cloned into pcDNA3.1+ N-terminal 3x-hemagglutinin tagged human Rho family small (Invitrogen) at KpnI (5') and XhoI (3'). The T19N mutation was GTP binding protein Rac3 (T17N mutant) cloned into pcDNA3.1+ introduced into the Rho family small GTP binding protein RhoA (Invitrogen) at KpnI (5') and XhoI (3'). The T17N mutation was (RHO0A00000) via the Quickchange mutagenesis kit introduced into Rac3 3X HA tagged (N-terminus)(RAC030TN00) (Stratagene). The mutation confers a dominant negative via the Quickchange mutagenesis kit (Stratagene). The mutation phenotype. Insert size = 680 bp. confers a dominant negative phenotype. Insert size = 670 bp.

RHO0A0TND0 RhoA T19N 3xHA-tagged(N-terminus) 2XMYC-tagged N-terminal 3x-hemagglutinin tagged human Rho family small

GTP binding protein RhoA (T19N mutant) cloned into RAC010MN00 Rac1 2XMYC-tagged(N-terminus) pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The T19N N-terminal 2XMYC-tagged human Rho family small GTP binding mutation was introduced into RhoA 3X HA tagged (N- protein Rac1 (wild type) cloned into pcDNA3.1+ (Invitrogen) at terminus)(RHO0A0TN00) via the Quickchange mutagenesis kit KpnI (5') and XhoI (3'). The open reading frame was derived (Stratagene). The mutation confers a dominant negative from RAC0100000 by the PCR. Insert size = 720 bp. phenotype. Insert size = 690 bp.

RHO0A0MN00 RhoA 2XMYC-tagged (N-terminus) RHO0B0TND0 RhoB T19N 3xHA-tagged(N-terminus) N-terminal 2XMYC-tagged human Rho family small GTP binding N-terminal 3x-hemagglutinin tagged human Rho family small protein RhoA (wild type) cloned into pcDNA3.1+ (Invitrogen) at GTP binding protein RhoB (T19N mutant) cloned into KpnI (5') and XhoI (3'). The open reading frame was derived pcDNA3.1+ (Invitrogen) at KpnI (5') and XbaI (3'). The T19N from RHO0A00000 by the PCR. Insert size = 680 bp. mutation was introduced into RhoB 3X HA tagged (N- terminus)(RHO0B0TN00) via the Quickchange mutagenesis kit

(Stratagene). The mutation confers a dominant negative phenotype. Insert size = 690 bp. GV PBRQ 2XMYC

RHO0C0TND0 RhoC T19N 3xHA-tagged(N-terminus) RHO0A0MNE0 RhoA G14V PBRQ 2XMYC-tagged (N- N-terminal 3x-hemagglutinin tagged human Rho family small terminus) GTP binding protein RhoC (T19N mutant) cloned into N-terminal 2XMYC-tagged human Rho family small GTP binding pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The T19N protein RhoA G14V (constitutively active) PBRQ (polybasic mutation was introduced into RhoC 3X HA tagged (N- region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') terminus)(RHO0C0TN00) via the Quickchange mutagenesis kit and XhoI (3'). The constitutively active and polybasic region (Stratagene). The mutation confers a dominant negative mutations were introduced into the Rho family small GTP binding phenotype. Insert size = 690 bp. protein RhoA (RHO0A00000) via Quickchange mutagenesis kit. (Stratagene). Phenotypes of these mutation are described in Lanning, Ruiz- Velasco and Williams, (2003) J. Biol. Chem. 2XMYC GV 278:12495-12506. Insert size = 680 bp.

RAC010MNC0 Rac1 G12V 2XMYC-tagged (N-terminus) GV PBRQ 3XHA N-terminal 2XMYC-tagged human Rho family small GTP binding protein Rac1 G12V (constitutively active) cloned into pcDNA3.1+ RAC010TNE0 Rac1 G12V PBRQ 3XHA-tagged (N-terminus) (Invitrogen) at KpnI (5') and XhoI (3'). The G12V mutation was N-terminal 3Xhemagglutinin-tagged human Rho family small introduced into the Rho family small GTP binding protein Rac1 GTP binding protein Rac1 G12V (constitutively active) PBRQ (RAC0100000) via the Quickchange mutagenesis kit (polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at (Stratagene). The mutation confers a constitutively active KpnI (5') and XhoI (3'). The G12V mutation and the PBRQ phenotype. Insert size = 720 bp. mutations were introduced into the Rho family small GTP binding protein Rac1 (RAC0100000) via the Quickchange mutagenesis RHO0A0MNC0 RhoA G14V 2XMYC-tagged(N-terminus) kit (Stratagene). The mutations confer a constitutively active and N-terminal 2XMYC-tagged human Rho family small GTP binding polybasic region mutated phenotype as described in Lanning, protein RhoA G14V (constitutively active) cloned into pcDNA3.1+ Ruiz-Velasco and Williams, (2003) J. Biol. Chem. 278:12495- (Invitrogen) at KpnI (5') and XhoI (3'). The G14V mutation was 12506. Insert size = 720 bp.

55 cDNA Resource Center

RHO0A0TNE0 RhoA G14V PBRQ 3XHA-tagged (N-terminus) TN PBRQ 3XHA N-terminal 3XHA-tagged human Rho family small GTP binding protein RhoA G14V (constitutively active) PBRQ (polybasic RAC010TNF0 Rac1 T17N PBRQ 3XHA-tagged (N-terminus) region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') N-terminal 3Xhemagglutinin-tagged human Rho family small and XhoI (3'). The G14V and polybasic region mutations were GTP binding protein Rac1 T17N (dominant negative) PBRQ introduced into the Rho family small GTP binding protein RhoA (polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at (RHO0A00000) via Quickchange mutagenesis kit (Stratagene). KpnI (5') and XhoI (3'). The T17N mutation and the PBRQ Phenotypes of these mutation are described in Lanning, Ruiz- mutations were introduced into the Rho family small GTP binding Velasco and Williams, (2003) J. Biol. Chem. 278:12495-12506. protein Rac1 (RAC0100000) via the Quickchange mutagenesis Insert size = 680 bp. kit (Stratagene). The mutations confer a dominant negative and polybasic region mutated phenotype as described in Lanning, Ruiz-Velasco and Williams, (2003) J. Biol. Chem. 278:12495- PBRQ 2XMYC 12506. Insert size = 720 bp.

RHO0A0MNB0 RhoA PBRQ 2XMYC-tagged (N-terminus) RHO0A0TNF0 RhoA T19N PBRQ 3XHA-tagged (N-terminus) N-terminal 2XMYC-tagged human Rho family small GTP binding N-terminal 3XHA-tagged human Rho family small GTP binding protein RhoA PBRQ (polybasic region mutated) cloned into protein RhoA T19N (dominant negative) PBRQ (polybasic pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') region mutations were introduced into the Rho family small GTP and XhoI (3'). The T19N and polybasic region mutations were binding protein RhoA (RHO0A00000) via Quickchange introduced into the Rho family small GTP binding protein RhoA mutagenesis kit (Stratagene). Phenotypes of these mutation are (RHO0A00000) via Quickchange mutagenesis kit (Stratagene). described in Lanning, Ruiz-Velasco and Williams, (2003) J. Biol. Phenotypes of these mutation are described in Lanning, Ruiz- Chem. 278:12495-12506. Insert size = 680 bp. Velasco and Williams, (2003) J. Biol. Chem. 278:12495-12506. Insert size = 680 bp.

PBRQ 3XHA

RAC010TNB0 Rac1 PBRQ 3xHA-tagged(N-terminus) N-terminal 3X-hemagglutinin tagged human Rho family small GTP binding protein Rac1 PBRQ (polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The polybasic region mutations were introduced into the Rho family small GTP binding protein Rac 1 (RAC0100000) via Quickchange mutagenesis kit (Stratagene). Phenotypes of these mutation are described in Lanning, Ruiz- Velasco and Williams, (2003) J. Biol. Chem. 278:12495-12506. Insert size = 730 bp.

RHO0A0TNB0 RhoA PBRQ 3XHA-tagged (N-terminus) N-terminal 3XHA-tagged human Rho family small GTP binding protein RhoA PBRQ (polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The polybasic region mutations were introduced into the Rho family small GTP binding protein RhoA (RHO0A00000) via Quickchange mutagenesis kit (Stratagene). Phenotypes of these mutation are described in Lanning, Ruiz-Velasco and Williams, (2003) J. Biol. Chem. 278:12495-12506. Insert size = 680 bp.

TN PBRQ 2XMYC

RHO0A0MNF0 RhoA T19N PBRQ 2XMYC-tagged (N- terminus) N-terminal 2XMYC-tagged human Rho family small GTP binding protein RhoA T19N (dominant negative) PBRQ (polybasic region mutated) cloned into pcDNA3.1+ (Invitrogen) at KpnI (5') and XhoI (3'). The T19N and polybasic region mutations were introduced into the Rho family small GTP binding protein RhoA (RHO0A00000) via Quickchange mutagenesis kit (Stratagene). Phenotypes of these mutation are described in Lanning, Ruiz- Velasco and Williams, (2003) J. Biol. Chem. 278:12495-12506. Insert size = 680 bp.

56 cDNA Resource Center

57