Product Information Sheet for NR-19667

Mycobacterium tuberculosis Gateway® Packaging/Storage: Clone Set, Recombinant in Escherichia NR-19667 was packaged aseptically in a 96-well plate. The product is provided frozen and should be stored at -80°C or coli, Plate 31 colder immediately upon arrival. For long-term storage, the vapor phase of a liquid nitrogen freezer is recommended. Catalog No. NR-19667 Freeze-thaw cycles should be avoided. This reagent is the tangible property of the U.S. Government. Growth Conditions: For research use only. Not for human use. Media: LB broth or agar containing 50 µg/mL kanamycin Incubation: Contributor: Temperature: 37°C Pathogen Functional Genomics Resource Center at the J. Atmosphere: Aerobic Craig Venter Institute Propagation:

1. Scrape top of frozen well with a pipette tip and streak onto Manufacturer: agar plate. BEI Resources 2. Incubate the plates at 37°C for 18 to 24 hours.

Product Description: Citation: Production in the 96-well format has increased risk of cross- Acknowledgment for publications should read “The following contamination between adjacent wells. Individual clones reagent was obtained through BEI Resources, NIAID, NIH: should be purified (e.g. single colony isolation and purification Mycobacterium tuberculosis Gateway® Clone Set, using good microbiological practices) and sequence-verified Recombinant in Escherichia coli, Plate 31, NR-19667.” prior to use. BEI Resources does not confirm or validate individual mutants provided by the contributor. Biosafety Level: 1

Appropriate safety procedures should always be used with this The Mycobacterium tuberculosis (M. tuberculosis), Gateway® material. Laboratory safety is discussed in the following clone set consists of 42 plates which contain 3724 sequence publication: U.S. Department of Health and Human Services, validated clones (3294 M. tuberculosis, strain H37Rv clones Public Health Service, Centers for Disease Control and supplemented with 430 unique open reading frames (ORF) Prevention, and National Institutes of Health. Biosafety in from M. tuberculosis, strain CDC1551) cloned in Escherichia Microbiological and Biomedical Laboratories. 5th ed. coli (E. coli) DH10B-T1 cells. Each ORF was recombined in Washington, DC: U.S. Government Printing Office, 2009; see vector pDONR221 with an ATG start codon and no stop www.cdc.gov/biosafety/publications/bmbl5/index.htm. codon. The sequence was validated by full length sequencing of each entry clone with greater than 1X coverage and a Disclaimers: mutation rate of less than 0.2%. Detailed information about You are authorized to use this product for research use only. each clone is shown in Table 1. It is not intended for human use.

Information related to the use of Gateway® Clones can be Use of this product is subject to the terms and conditions of obtained from Invitrogen. Recombination was facilitated the BEI Resources Material Transfer Agreement (MTA). The through an attB substrate (attB-PCR product or a linearized MTA is available on our Web site at www.beiresources.org. attB expression clone) with an attP substrate (pDONR221) to create an attL-containing entry clone. The entry clone While BEI Resources uses reasonable efforts to include contains recombinational cloning sites, attL1 and attL2 to accurate and up-to-date information on this product sheet, facilitate gene transfer into a destination vector, M13 forward neither ATCC® nor the U.S. Government makes any and reverse priming sites for sequencing and a kanamycin warranties or representations as to its accuracy. Citations resistance gene for selection. Please refer to the Invitrogen from scientific literature and patents are provided for ® Gateway Technology Manual for additional details. informational purposes only. Neither ATCC® nor the U.S. Government warrants that such information has been Plate orientation and viability were confirmed for NR-19667. confirmed to be accurate.

Material Provided: This product is sent with the condition that you are responsible Each inoculated well of the 96-well plate contains for its safe storage, handling, use and disposal. ATCC® and approximately 60 µL of culture in Luria Bertani (LB) broth the U.S. Government are not liable for any damages or injuries containing 50 µg/mL kanamycin supplemented with 15% arising from receipt and/or use of this product. While glycerol. reasonable effort is made to ensure authenticity and reliability of materials on deposit, the U.S. Government, ATCC®, their suppliers and contributors to BEI Resources are not liable for

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Product Information Sheet for NR-19667

damages arising from the misidentification or References: misrepresentation of products. 1. Cole, S. T., et al. “Deciphering the Biology of Mycobacterium tuberculosis from the Complete Genome Use Restrictions: Sequence.” Nature 393 (1998): 537-544. PubMed: This material is distributed for internal research, non- 9634230. commercial purposes only. This material, its product or its 2. Camus, J. C., et al. “Re-Annotation of the Genome derivatives may not be distributed to third parties. Except as Sequence of Mycobacterium tuberculosis H37Rv.” performed under a U.S. Government contract, individuals Microbiology 148 (2002): 2967-2973. PubMed: contemplating commercial use of the material, its products or 12368430. its derivatives must contact the contributor to determine if a license is required. U.S. Government contractors may need a ATCC® is a trademark of the American Type Culture license before first commercial sale. Collection.

Table 1: Mycobacterium tuberculosis, Gateway® Clones, Plate 31 (ZMTLU)1 Clone Well ORF Locus ID Description (Gene name) Accession Average Depth Position Length Number of Coverage 43313 A01 1111 Rv1112 GTP-dependent nucleic acid-binding protein EngD NP_215628.1 4.720072007 43320 A02 1111 Rv3497c MCE-family protein MCE4C NP_218014.1 4.747074707 43314 A03 1111 Rv1299 peptide chain release factor 1 NP_215815.1 3.936993699 43324 A04 1114 Rv0926c hypothetical protein Rv0926c NP_215441.1 2.859964093 43331 A05 1114 Rv3571 hemoglobine-like protein NP_218088.1 3.961400359 43336 A06 1114 Rv3731 ATP-dependent DNA ligase NP_218248.1 3.946140036 43327 A07 1114 Rv2138 lipoprotein LppL NP_216654.1 4.772890485 43330 A09 1114 Rv3037c hypothetical protein Rv3037c NP_217553.1 4.438061041 43322 A10 1114 Rv0502 hypothetical protein Rv0502 NP_215016.1 4.745960503 43338 A11 1117 Rv0855 fatty-acid-CoA racemase NP_215370.1 4.692927484 D-alpha-D-mannose-1-phosphate 43346 A12 1117 Rv3264c guanylyltransferase MANB (D-alpha-D-heptose-1- YP_177951.1 4.642793196 phosphate guanylyltransferase) 43337 B01 1117 Rv0655 ribonucleotide ABC transporter ATP-binding protein NP_215169.1 4.736794987 43345 B02 1117 Rv3109 molybdenum cofactor biosynthesis protein A YP_177925.1 4.000895255 multifunctional dimethylallyltransferase/farnesyl 43347 B03 1117 Rv3398c YP_177970.1 4.742166517 diphosphate synthetase/ farnesyltranstransferase 43356 B04 1120 Rv1295 threonine synthase NP_215811.1 4.733928571 43354 B05 1120 Rv1143 alpha-methylacyl-CoA racemase NP_215659.1 2.122321429 43357 B06 1120 Rv1607 ionic transporter integral membrane protein chaA NP_216123.1 3.132142857 43353 B07 1120 Rv1063c hypothetical protein Rv1063c NP_215579.1 3.86875 putative f420-dependent glucose-6-phosphate 43350 B08 1120 Rv0132c NP_214646.1 4.11875 dehydrogenase Fgd2 sn-glycerol-3-phosphate transport ATP-binding 43359 B09 1120 Rv2832c NP_217348.1 4.727678571 protein ABC transporter UGPC 43358 B10 1120 Rv2262c hypothetical protein Rv2262c NP_216778.1 4.73125 nicotinate-nucleotide--dimethylbenzimidazole 43363 B11 1123 Rv2207 NP_216723.1 4.750667854 phosphoribosyltransferase 43362 B12 1123 Rv0209 hypothetical protein Rv0209 NP_214723.1 3.960819234 43374 C01 1126 Rv3138 pyruvate formate lyase activating protein PflA NP_217654.1 4.431616341 43370 C02 1126 Rv1009 resuscitation-promoting factor rpfB NP_215525.1 4.719360568 43372 C03 1126 Rv1178 N-succinyldiaminopimelate aminotransferase NP_215694.1 4.414742451 43373 C04 1126 Rv2538c 3-dehydroquinate synthase NP_217054.1 4.379218472 43379 C05 1129 Rv2064 cobalamin biosynthesis protein CobG NP_216580.1 4.715677591 transmembrane cytochrome C oxidase subunit II 43380 C06 1129 Rv2200c NP_216716.1 4.389725421 CtaC 43377 C07 1129 Rv1933c acyl-CoA dehydrogenase FADE18 NP_216449.1 4.729849424 43384 C08 1132 Rv0809 phosphoribosylaminoimidazole synthetase NP_215324.1 4.71819788 43383 C09 1132 Rv0797 IS1547 transposase NP_215312.1 2.479681979

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Product Information Sheet for NR-19667

Clone Well ORF Locus ID Description (Gene name) Accession Average Depth Position Length Number of Coverage type I restriction/modification system specificity 43388 C10 1132 Rv2761c NP_217277.1 4.696996466 determinant HsdS 43385 C11 1132 Rv2228c bifunctional RNase H/acid phosphatase NP_216744.1 4.720848057 43395 C12 1135 Rv3313c adenosine deaminase NP_217830.1 3.856387665 43391 D01 1135 Rv0029 hypothetical protein Rv0029 NP_214543.1 4.672246696 43392 D02 1135 Rv1765c transposase YP_177652.1 4.626431718 43397 D03 1135 Rv3629c hypothetical protein Rv3629c NP_218146.1 4.678414097 43399 D04 1138 Rv0192 hypothetical protein Rv0192 NP_214706.1 4.699472759 43401 D05 1138 Rv1239c magnesium/cobalt transporter CorA NP_215755.1 4.731107206 43398 D06 1138 Rv0155 NAD(P) transhydrogenase subunit alpha NP_214669.1 4.69771529 43415 D07 1141 Rv3024c tRNA-specific 2-thiouridylase MnmA NP_217540.1 2.914110429 43410 D08 1141 Rv1915 isocitrate lyase NP_216431.1 4.695004382 43408 D09 1141 Rv0726c hypothetical protein Rv0726c NP_215240.1 4.695880806 myo-inositol-1-phosphate synthase INO1 (inositol 1- phosphate synthetase) (D-glucose 6-phosphate 43405 D10 1141 Rv0046c NP_214560.1 2.406660824 cycloaldolase) (glucose 6-phosphate cyclase) (glucocycloaldolase) pyruvate dehydrogenase E1 component alpha 43413 D11 1141 Rv2497c NP_217013.1 2.412795793 subunit PdhA 43409 D12 1141 Rv1530 alcohol dehydrogenase adh NP_216046.1 3.586327783 43416 E01 1144 Rv0811c hypothetical protein Rv0811c NP_215326.1 4.666958042 43418 E02 1144 Rv2210c branched-chain amino acid aminotransferase NP_216726.1 3.19493007 43421 E03 1144 Rv3873 PPE family protein YP_178022.1 4.692307692 43426 E04 1147 Rv1859 molybdenum ABC transporter ATP-binding protein NP_216375.1 3.934612031 43430 E05 1147 Rv2670c hypothetical protein Rv2670c NP_217186.1 2.66695728 43425 E06 1147 Rv0482 UDP-N-acetylenolpyruvoylglucosamine reductase NP_214996.1 4.667829119 43423 E07 1147 Rv0179c lipoprotein LprO NP_214693.1 4.669572799 43431 E08 1147 Rv3071 hypothetical protein Rv3071 NP_217587.1 4.316477768 43439 E09 1150 Rv2915c hypothetical protein Rv2915c NP_217431.1 4.606086957 43438 E10 1150 Rv2192c anthranilate phosphoribosyltransferase NP_216708.1 3.524347826 periplasmic phosphate-binding lipoprotein PSTS2 43434 E11 1150 Rv0932c YP_177769.1 4.335652174 (PBP-2) (PSTS2) 43435 E12 1150 Rv1106c cholesterol dehydrogenase NP_215622.1 3.897391304 43432 F01 1150 Rv0182c RNA polymerase factor sigma-70 NP_214696.1 4.675652174 periplasmic phosphate-binding lipoprotein PSTS3 43433 F02 1150 Rv0928 YP_177768.1 3.991304348 (PBP-3) (PSTS3) (PHOS1) secreted L-alanine dehydrogenase ALD (40 kDa 43445 F03 1153 Rv2780 NP_217296.1 4.671292281 antigen) (TB43) 43441 F04 1153 Rv1048c hypothetical protein Rv1048c NP_215564.1 4.13876843 43444 F05 1153 Rv1922 lipoprotein NP_216438.1 4.522983521 macrolide-transport ATP-binding protein ABC 43450 F06 1156 Rv1668c NP_216184.1 4.621107266 transporter 43448 F07 1156 Rv0906 hypothetical protein Rv0906 NP_215421.1 4.288062284 43446 F09 1156 Rv0848 cysteine synthase A CysK2 YP_177762.1 4.666089965 43451 F10 1156 Rv2337c hypothetical protein Rv2337c NP_216853.1 4.393598616 43456 F11 1156 Rv3215 isochorismate synthase NP_217731.1 4.686851211 43449 F12 1156 Rv1260 hypothetical protein Rv1260 NP_215776.1 4.670415225 43462 G01 1159 Rv1679 acyl-CoA dehydrogenase NP_216195.1 4.664365833 43459 G02 1159 Rv1448c transaldolase NP_215964.1 3.868852459 43457 G03 1159 Rv0505c phosphoserine phosphatase YP_177732.1 4.331320104 43458 G04 1159 Rv0889c citrate synthase 2 NP_215404.1 4.581535807 43466 G05 1159 Rv2981c D-alanyl-alanine synthetase A NP_217497.1 2.584987058 43468 G06 1159 Rv3505 acyl-CoA dehydrogenase FADE27 NP_218022.1 4.661777394 43467 G07 1159 Rv3006 lipoprotein LppZ NP_217522.1 4.686798965

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Product Information Sheet for NR-19667

Clone Well ORF Locus ID Description (Gene name) Accession Average Depth Position Length Number of Coverage 2-oxoglutarate ferredoxin oxidoreductase subunit 43463 G08 1159 Rv2454c NP_216970.1 4.681622088 beta 43470 G09 1162 Rv2629 hypothetical protein Rv2629 NP_217145.1 4.683304647 43471 G10 1162 Rv3169 hypothetical protein Rv3169 NP_217685.1 4.674698795 periplasmic phosphate-binding lipoprotein PSTS1 43469 G11 1162 Rv0934 YP_177770.1 4.669535284 (PBP-1) (PSTS1) 43477 G12 1165 Rv2388c coproporphyrinogen III oxidase NP_216904.1 3.164806867 43484 H02 1165 Rv3531c hypothetical protein Rv3531c NP_218048.1 3.065236052 43485 H03 1165 Rv3796 hypothetical protein Rv3796 YP_178016.1 3.644635193 zinc-containing alcohol dehydrogenase NAD 43472 H04 1165 Rv0761c YP_177754.1 4.497854077 dependent ADHB 43480 H05 1165 Rv2819c hypothetical protein Rv2819c NP_217335.1 4.150214592 43478 H06 1165 Rv2659c phiRv2 prophage integrase NP_217175.1 4.652360515 43473 H07 1165 Rv1533 hypothetical protein Rv1533 NP_216049.1 4.681545064 43497 H08 1168 Rv2970c hypothetical protein Rv2970A YP_177681.1 3.988013699 43498 H09 1168 Rv3094c hypothetical protein Rv3094c NP_217610.1 4.667808219 43486 H10 1168 Rv0365c hypothetical protein Rv0365c NP_214879.1 4.080479452 43494 H11 1168 Rv1894c hypothetical protein Rv1894c NP_216410.1 3.659246575 43491 H12 1168 Rv0884c phosphoserine aminotransferase NP_215399.1 4.659246575 1All information in this table was provided by J. Craig Venter Institute at the time of deposition.

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