OMP Nucleotide Sequences

OMP nucleotide sequences Vector: pET11a OmpX

Mature OmpX Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGGCGACTTCTACTGTAACTGGCGGT TACGCACAGAGCGACGCTCAGGGCCAAATGAACAAAATGGGCGGTTTCAACCTGAA ATACCGCTATGAAGAAGACAACAGCCCGCTGGGTGTGATCGGTTCTTTCACTTACAC CGAGAAAAGCCGTACTGCAAGCTCTGGTGACTACAACAAAAACCAGTACTACGGCA TCACTGCTGGTCCGGCTTACCGCATTAACGACTGGGCAAGCATCTACGGTGTAGTGG GTGTGGGTTATGGTAAATTCCAGACCACTGAATACCCGACCTACAAACACGACACCA GCGACTACGGTTTCTCCTACGGTGCGGGTCTGCAGTTCAACCCGATGGAAAACGTTG CTCTGGACTTCTCTTACGAGCAGAGCCGTATTCGTAGCGTTGACGTAGGCACCTGGAT TGCCGGTGTTGGTTACCGCTTCTAAGGATCCGGCTGCTAACAAAGCCCGAAAGGAAG CTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTA AACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGAT

Cloning details: Cloned by Thuy

Forward primer: 5’ GCTA CATATG GCG ACT TCT ACT GTA ACT GGC 3’

Reverse complement of reverse primer: 5’ GGT GTT GGT TAC CGC TTC TAA GGATCC GCC 3’

1 PagP

Mature PagP Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGAACGCAGATGAGTGGATGACAAC GTTTAGAGAAAATATTGCACAAACCTGGCAACAGCCTGAACATTATGATTTATATAT TCCTGCCATCACCTGGCATGCACGTTTCGCTTACGACAAAGAAAAAACCGATCGCTA TAACGAGCGACCGTGGGGTGGCGGTTTTGGCCTGTCGCGTTGGGATGAAAAAGGAA ACTGGCATGGCCTGTATGCCATGGCATTTAAGGACTCGTGGAACAAATGGGAACCGA TTGCCGGATACGGATGGGAAAGTACCTGGCGACCGCTGGCGGATGAAAATTTTCATT TAGGTCTGGGATTCACCGCTGGCGTAACGGCACGCGATAACTGGAATTACATCCCTC TCCCGGTTCTACTGCCATTGGCCTCCGTGGGTTATGGCCCAGTGACTTTTCAGATGAC CTACATTCCGGGTACCTACAACAATGGCAATGTGTACTTTGCCTGGATGCGCTTTCAG TTTTGAGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCAC CGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTTAGGGGTTT TTTGCTGAAAGGAGGAACTATATCCGGAT

Forward primer: 5’ GCTA CATATG AAC GCA GAT GAG TGG ATG AC 3’

Reverse complement of reverse primer: 5’ GG ATG CGC TTT CAG TTT TGA GGATCC GCC 3’

2 OmpW

Mature OmpW Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGCATGAAGCAGGCGAATTTTTTATG CGTGCAGGTTCTGCAACCGTACGTCCAACAGAAGGTGCTGGTGGTACGTTAGGAAGT CTGGGTGGATTCAGCGTGACCAATAACACGCAACTGGGCCTTACGTTTACTTATATG GCGACCGACAACATTGGTGTGGAATTACTGGCAGCGACGCCGTTCCGCCATAAAATC GGCACCCGGGCGACCGGCGATATTGCAACCGTTCATCATCTGCCACCAACACTGATG GCGCAGTGGTATTTTGGTGATGCCAGCAGCAAATTCCGTCCTTACGTTGGGGCAGGT ATTAACTACACCACCTTCTTTGATAATGGATTTAACGATCATGGCAAAGAGGCAGGG CTTTCCGATCTCAGTCTGAAAGATTCCTGGGGAGCTGCCGGGCAGGTGGGGGTTGAT TATCTGATTAACCGTGACTGGTTGGTTAACATGTCAGTGTGGTACATGGATATCGATA CCACCGCCAATTATAAGCTGGGCGGTGCACAGCAACACGATAGCGTACGCCTCGATC CGTGGGTGTTTATGTTCTCAGCAGGATATCGTTTTTAAGGATCCGGCTGCTAACAAAG CCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCC TTGGGGCCTCTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCG GAT

Forward primer: 5’ GCTA CATATG CAT GAA GCA GGC GAA TTT TTT ATG 3’

Reverse complement of reverse primer: 5’ G TTC TCA GCA GGA TAT CGT TTT TAA GGATCC GCC 3’

3 OmpA

Mature OmpA (has mutation Nancy made to remove internal BamHI site) Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGGCTCCGAAAGATAACACCTGGTA CACTGGTGCTAAACTGGGCTGGTCCCAGTACCATGACACTGGTTTCATCAACAACAA TGGCCCGACCCATGAAAACCAACTGGGCGCTGGTGCTTTTGGTGGTTACCAGGTTAA CCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTAGGTCGTATGCCGTACAAAGG CAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGG TTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCG TGCAGACACTAAATCCAACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGT CTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGTCTGGAATAC CAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGG CATGCTGAGCCTGGGTGTTTCCTACCGTTTCGGTCAGGGCGAAGCAGCTCCAGTAGTT GCTCCGGCTCCAGCTCCGGCACCGGAAGTACAGACCAAGCACTTCACTCTGAAGTCT GACGTTCTGTTCAACTTCAACAAAGCAACCCTGAAACCGGAAGGTCAGGCTGCTCTG GATCAGCTGTACAGCCAGCTGAGCAACCTGGACCCGAAAGACGGTTCCGTAGTTGTT CTGGGTTACACCGACCGCATCGGTTCTGACGCTTACAACCAGGGTCTGTCCGAGCGC CGTGCTCAGTCTGTTGTTGATTACCTGATCTCCAAAGGTATCCCGGCAGACAAGATCT CCGCACGTGGTATGGGCGAATCCAACCCGGTTACTGGCAACACCTGTGACAACGTGA AACAGCGTGCTGCACTGATCGACTGCCTGGCTCCGGATCGTCGCGTAGAGATCGAAG TTAAAGGTATCAAAGACGTTGTAACTCAGCCGCAGGCTTAAGGATCCGGCTGCTAAC AAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATA ACCCCTTGGGGCCTCTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTAT ATCCGGAT

One mutation: GAC instead of GAT (both code for Asp)

Cloning details: Cloned by Nancy as “OmpA326”

Forward primer used: 5’ GGCATCC CATATG GCT CCG AAA GAT AAC ACC TG 3’

Reverse complement of reverse primer used: 5’ GTA ACT CAG CCG CAG GCT TAA GGATCC CG 3’

4 OmpA171

Mature OmpA171 (OmpA barrel only) Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGGCTCCGAAAGATAACACCTGGTA CACTGGTGCTAAACTGGGCTGGTCCCAGTACCATGACACTGGTTTCATCAACAACAA TGGCCCGACCCATGAAAACCAACTGGGCGCTGGTGCTTTTGGTGGTTACCAGGTTAA CCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTAGGTCGTATGCCGTACAAAGG CAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGG TTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCG TGCAGACACTAAATCCAACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGT CTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGTCTGGAATAC CAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGG CATGCTGAGCCTGGGTGTTTCCTACCGTTTTGGTTGAGGATCCGGCTGCTAACAAAGC CCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCT TGGGGCCTCTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGG AT

One mutation: TTT instead of TTC (both code for Phe).

Cloning details: Cloned by Nancy as “OmpA171” using primers from Pautsch, et al. Proteins 1999; 34: 167-172.

Forward primer used: 5’ GGCATCC CATATG GCT CCG AAA GAT AAC ACC TG 3’

Reverse complement of reverse primer used: 5’ GGT GTT TCC TAC CGT TTT GGT TGA GGATCC CG 3’

The mutation is in Pautsch’s reverse primer so they must have chosen to make the mutation for reasons unknown to me (I verified this by looking up the paper but they don’t say why). It doesn’t change the amino acid though so Nancy went with it.

5 OmpT

Mature OmpT Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGTCTACCGAGACTTTATCGTTTACT CCTGACAACATAAATGCGGACATTAGTCTTGGAACTCTGAGCGGAAAAACAAAAGA GCGTGTTTATCTAGCCGAAGAAGGAGGCCGAAAAGTCAGTCAACTCGACTGGAAATT CAATAACGCTGCAATTATTAAAGGTGCAATTAATTGGGATTTGATGCCCCAGATATC TATCGGGGCTGCTGGCTGGACAACTCTCGGCAGCCGAGGTGGCAATATGGTCGATCA GGACTGGATGGATTCCAGTAACCCCGGAACCTGGACGGATGAAAGTAGACACCCTG ATACACAACTCAATTATGCCAACGAATTTGATCTGAATATCAAAGGCTGGCTCCTCA ACGAACCCAATTACCGCCTGGGACTCATGGCCGGATATCAGGAAAGCCGTTATAGCT TTACAGCCAGAGGTGGTTCCTATATCTACAGTTCTGAGGAGGGATTCAGAGATGATA TCGGCTCCTTCCCGAATGGAGAAAGAGCAATCGGCTACAAACAACGTTTTAAAATGC CCTACATTGGCTTGACTGGAAGTTATCGTTATGAAGATTTTGAACTCGGTGGCACATT TAAATACAGCGGCTGGGTGGAATCATCTGATAACGATGAACACTATGACCCGGGAA AAAGAATCACTTATCGCAGTAAGGTCAAAGACCAAAATTACTATTCTGTTGCAGTCA ATGCAGGTTATTACGTCACACCTAACGCAAAAGTTTATGTTGAAGGCGCATGGAATC GGGTTACGAATAAAAAAGGTAATACTTCACTTTATGATCACAATAATAACACTTCAG ACTACAGCAAAAATGGAGCAGGTATAGAAAACTATAACTTCATCACTACTGCTGGTC TTAAGTACACATTTTAAGGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTG GCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTC TTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGAT

Forward primer: 5’ GCTA CATATG TCT ACC GAG ACT TTA TCG TTT ACT CC 3’

Reverse complement of reverse primer: 5’ CT GCT GGT CTT AAG TAC ACA TTT TAA GGATCC GCC 3’

6 OmpLa

Mature OmpLa Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGCAAGAGGCAACGGTGAAAGAGGT GCATGACGCGCCAGCGGTGCGTGGCAGTATTATCGCCAATATGCTGCAGGAGCATGA CAATCCGTTCACGCTCTATCCTTATGACACCAACTACCTCATTTACACCCAAACCAGC GATCTGAATAAAGAAGCGATTGCCAGTTACGACTGGGCGGAAAATGCGCGTAAGGA TGAAGTAAAGTTTCAGTTGAGCCTGGCATTTCCGCTGTGGCGTGGGATTTTAGGCCCG AACTCGGTGTTGGGTGCGTCTTATACGCAAAAATCCTGGTGGCAACTGTCCAATAGC GAAGAGTCTTCACCGTTTCGTGAAACCAACTACGAACCGCAATTGTTCCTCGGTTTTG CCACCGATTACCGTTTTGCAGGTTGGACGCTGCGCGATGTGGAGATGGGGTATAACC ACGACTCTAACGGGCGTTCCGACCCGACCTCCCGCAGCTGGAACCGCCTTTATACTC GCCTGATGGCAGAAAACGGTAACTGGCTGGTAGAAGTGAAGCCGTGGTATGTGGTG GGTAATACTGACGATAACCCGGATATCACCAAATATATGGGTTACTACCAGCTTAAA ATCGGCTATCACCTCGGCGATGCGGTGCTCAGTGCGAAAGGACAGTACAACTGGAAC ACCGGCTACGGCGGCGCGGAGTTAGGCTTAAGTTACCCGATCACCAAACATGTGCGC CTTTATACTCAGGTTTACAGCGGCTATGGCGAATCGCTCATCGACTATAACTTCAACC AGACCCGTGTCGGTGTGGGGGTTATGCTAAACGATTTGTTTTAAGAATGCGGATCCG GCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATA ACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGG AGGAACTATATCCGGAT

One mutation: GGC instead of GGT (both code for glycine) No idea if someone put this here on purpose or why.

Cloning details: (from Karen’s word document on Ompla primer design)

Forward primer used: 5’ GGCAGTA CATATG CAA GAG GCA ACG GTG AAA G 3’

Reverse complement of reverse primer used: 5’ G GTT ATG CTA AAC GAT TTG TTT TAA GAATGC GGATCC CG 3’

GAATGC is recognition site for BsmI for use during cloning (unique site to check if insert is present in plasmid).

7 FadL

Mature FadL Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGGCAGGCTTTCAGTTAAACGAATTT TCTTCCTCTGGCCTGGGCCGGGCTTATTCAGGGGAAGGCGCAATTGCCGATGATGCA GGTAACGTCAGCCGTAACCCCGCATTGATTACTATGTTTGACCGCCCGACATTTTCTG CGGGTGCGGTTTATATTGACCCGGATGTAAATATCAGCGGAACGTCTCCATCTGGTC GTAGCCTGAAAGCCGATAACATCGCGCCTACGGCATGGGTTCCGAACATGCACTTTG TTGCACCGATTAACGACCAATTTGGTTGGGGCGCTTCTATTACCTCTAACTATGGTCT GGCTACAGAGTTTAACGATACTTATGCAGGCGGCTCTGTCGGGGGTACAACCGACCT TGAAACCATGAACCTGAACTTAAGCGGTGCGTATCGCTTAAATAATGCATGGAGCTT TGGTCTTGGTTTCAACGCCGTCTACGCTCGCGCGAAAATTGAACGTTTCGCAGGCGAT CTGGGGCAGTTGGTTGCTGGCCAAATTATGCAATCTCCTGCTGGCCAAACTCAGCAA GGGCAAGCATTGGCAGCTACCGCCAACGGTATTGACAGTAATACCAAAATCGCTCAT CTGAACGGTAACCAGTGGGGCTTTGGCTGGAACGCCGGAATCCTGTATGAACTGGAT AAAAATAACCGCTATGCACTGACCTACCGTTCTGAAGTGAAAATTGACTTCAAAGGT AACTACAGCAGCGATCTTAATCGTGCGTTTAATAACTACGGTTTGCCAATTCCTACCG CGACAGGTGGCGCAACGCAATCGGGTTATCTGACGCTGAACCTGCCTGAAATGTGGG AAGTGTCAGGTTATAACCGTGTTGATCCACAGTGGGCGATTCACTATAGCCTGGCTT ACACCAGCTGGAGTCAGTTCCAGCAGCTGAAAGCGACCTCAACCAGTGGCGACACG CTGTTCCAGAAACATGAAGGCTTTAAAGATGCTTACCGCATCGCGTTGGGTACCACT TATTACTACGATGATAACTGGACCTTCCGTACCGGTATCGCCTTTGATGACAGCCCAG TTCCTGCACAGAATCGTTCTATCTCCATTCCGGACCAGGACCGTTTCTGGCTGAGTGC AGGTACGACTTACGCATTTAATAAAGATGCTTCAGTCGACGTTGGTGTTTCTTATATG CACGGTCAGAGCGTGAAAATTAACGAAGGCCCATACCAGTTCGAGTCTGAAGGTAA AGCCTGGCTGTTCGGTACTAACTTTAACTACGCGTTCTGAGGATCCGGCTGCTAACAA AGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACC CCTTGGGGCCTCTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCC GGAT

Forward primer: 5’ GCTA CATATG GCA GGC TTT CAG TTA AAC GAA TTT TC 3’

Reverse complement of reverse primer: 5’ GGT ACT AAC TTT AAC TAC GCG TTC TGA GGATCC GCC 3’

8 Omp85

Mature Omp85 (YaeT, BamA) Start and stop codons Restriction enzyme sites (CATATG=NdeI and GGATCC=BamHI) T7 Promoter with lac operator Ribosome binding site T7 terminator

TAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAA TTTTGTTTAACTTTAAGAAGGAGATATACATATGGAAGGGTTCGTAGTGAAAGATAT TCATTTCGAAGGCCTTCAGCGTGTCGCCGTTGGTGCGGCCCTCCTCAGTATGCCGGTG CGCACAGGCGACACGGTTAATGATGAAGATATCAGTAATACCATTCGCGCTCTGTTT GCTACCGGCAACTTTGAGGATGTTCGCGTCCTTCGTGATGGTGATACCCTTCTGGTTC AGGTAAAAGAACGTCCGACCATTGCCAGCATTACTTTCTCCGGTAACAAATCGGTGA AAGATGACATGCTGAAGCAAAACCTCGAGGCTTCTGGTGTGCGTGTGGGCGAATCCC TCGATCGCACCACCATTGCCGATATCGAGAAAGGTCTGGAAGACTTCTACTACAGCG TCGGTAAATATAGCGCCAGCGTAAAAGCTGTCGTGACCCCGCTGCCGCGCAACCGTG TTGACCTAAAACTGGTGTTCCAGGAAGGTGTGTCAGCTGAAATCCAGCAAATTAACA TTGTTGGTAACCATGCTTTCACCACCGACGAACTGATCTCTCATTTCCAACTGCGTGA CGAAGTGCCGTGGTGGAACGTGGTAGGCGATCGTAAATACCAGAAACAGAAACTGG CGGGCGACCTTGAAACCCTGCGCAGCTACTATCTGGATCGCGGTTATGCCCGTTTCA ACATCGACTCTACCCAGGTCAGTCTGACGCCAGATAAAAAAGGTATTTACGTCACGG TGAACATCACCGAAGGCGATCAGTACAAGCTTTCTGGCGTTGAAGTGAGCGGCAACC TTGCCGGGCACTCCGCTGAAATTGAGCAGCTGACTAAGATCGAGCCGGGTGAGCTGT ATAACGGCACCAAAGTGACCAAGATGGAAGATGACATCAAAAAGCTTCTCGGTCGC TATGGTTATGCCTATCCGCGCGTACAGTCGATGCCCGAAATTAACGATGCCGACAAA ACCGTTAAATTACGTGTGAACGTTGATGCGGGTAACCGTTTCTACGTGCGTAAGATC CGTTTTGAAGGTAACGATACCTCGAAAGATGCCGTCCTGCGTCGCGAAATGCGTCAG ATGGAAGGTGCATGGCTGGGGAGCGATCTGGTCGATCAGGGTAAGGAGCGTCTGAA TCGTCTGGGCTTCTTTGAAACTGTCGATACCGATACCCAACGTGTTCCGGGTAGCCCG GACCAGGTTGATGTCGTCTACAAGGTAAAAGAGCGCAACACCGGTAGCTTCAACTTT GGTATTGGTTACGGTACTGAAAGTGGCGTGAGCTTCCAGGCTGGTGTGCAGCAGGAT AACTGGTTAGGTACAGGTTATGCTGTTGGTATCAACGGGACCAAAAACGATTACCAG ACCTATGCTGAACTGTCGGTAACCAACCCGTACTTCACCGTAGATGGCGTAAGCCTC GGTGGTCGTCTCTTCTATAATGACTTCCAGGCAGATGACGCCGACCTGTCCGACTATA CCAACAAGAGTTATGGTACAGACGTGACGTTGGGCTTCCCGATTAACGAATATAACT CGCTGCGTGCAGGTCTGGGTTATGTACATAACTCCCTGTCCAACATGCAGCCTCAGGT TGCGATGTGGCGTTATCTGTACTCTATGGGTGAACATCCGAGCACCTCTGATCAGGAT AACAGCTTCAAAACGGACGACTTCACGTTCAACTATGGTTGGACCTATAACAAGCTT GACCGTGGTTACTTCCCGACAGATGGTTCACGTGTCAACCTGACCGGTAAAGTGACC ATTCCTGGATCGGATAACGAATACTACAAAGTGACGTTAGACACGGCGACTTATGTG CCGATCGATGACGATCACAAATGGGTTGTTCTGGGGCGTACCCGCTGGGGTTATGGT GATGGTTTAGGCGGCAAAGAGATGCCGTTCTACGAGAACTTCTATGCCGGTGGTTCC AGCACCGTGCGTGGCTTCCAGTCCAATACCATTGGTCCGAAAGCAGTTTACTTCCCGC ATCAGGCCAGTAATTATGATCCGGACTATGATTACGAATGTGCGACTCAGGACGGCG CGAAAGACCTGTGTAAATCGGATGATGCTGTAGGCGGTAACGCCATGGCGGTTGCCA GCCTCGAGTTCATCACCCCGACGCCGTTTATTAGCGATAAGTATGCTAACTCGGTTCG TACTTCCTTCTTCTGGGATATGGGTACCGTTTGGGATACAAACTGGGATTCCAGCCAA

9 TATTCTGGATATCCGGACTATAGTGATCCAAGCAATATCCGTATGTCTGCGGGTATCG CATTACAATGGATGTCCCCATTGGGGCCGTTGGTGTTCTCCTACGCCCAGCCGTTCAA AAAGTACGATGGAGACAAGGCAGAACAGTTCCAGTTTAACATCGGTAAAACCTGGT AAGTGTTCTCCACAAAGGAATGTAGTGGGATCCGGCTGCTAACAAAGCCCGAAAGG AAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCT CTAAACGGGTCTTTAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGAT

Cloning details: Re-cloned by Thuy (after Ann Marie’s first attempt)

Forward primer: 5’ GCTA CATATG GAA GGG TTC GTA GTG AAA GAT AT 3’

Reverse complement of reverse primer: 5’ GT TCT CCA CAA AGG AAT GTA GTG GGATCC GCC 3’

Thuy designed the reverse primer to clone beyond the stop codon because they were having problems with expression and Karen theorized that it might have to do with mRNA stability.