COMMENT pMCSG50 6347 bp BL21 BirA gene cloned into the SmaI LIC site of MCSG60. 15 aa biotinylation signal with 6aa His tag inserted between the NdeI and Acc65I sites. Reference for biotinylation sequence: Beckett et al. (1999) Protein Science 8:921-929 FEATURES Location/Qualifiers misc_feature complement (1232..1263) /note="MCSG7 LIC site" Cleavage-site complement (1251..1271) /note="TEV Site" Region complement (1329..1346) /note="His Tag" promoter complement(1421..1437) /note="T7 Promoter" terminator complement(26..72) /note="T7 Terminator" CDS 1951..2910 /note="Lac I cds" CDS complement(5099..5959) /note="bla cds" rep_origin complement (4340..4928) /note="pBR322 origin of replication" RBS complement(1215..1220) /note="ribosome binding site" RBS complement(1358..1363) /note="ribosome binding site" CDS complement(244..1206) /note="BirA cds" Binding-site 1278..1322 /note="biotinylation signal" 1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA 61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT 121 TGTTAGCAGC CGGATCTCAG TGGTGGTGGT GGTGGTGCTC GAGTGCGGCC GCAAGCTTGT 181 CGACGGAGCT CGAATTCGGA TCCGTTATCC ACTTCCAATG TTATGGTGGT GCCCAGCTTA 241 TTATTTTTCT GCACTACGCA GGGATATTTC ACCGCCCATC CAGGGTTTTA TTATTCCATC 301 CTGCTCAAGT AATAAAGCCC CCTGTTTGTC TATTCCGCGT GAAATGCCAA ATATCTCTTT 361 ATCACCAATG ATAAGTTTCA CTGGGCGATT AATAAAATTA TCCAGCTTTT CCCAGCGCGA 421 CAGATAAGGT GCCAATCCTT CTTGTTCGAA GAGTTCCAAC GCAGCACGTA ATTCACGTAT 481 TAGCATGGCC GCCAACGTAT TACGATCGAG ATTGATCCCC GCTTCCTGCA GCGTGATCCA 541 CCCCTGATTA ACGACACTCT CTTCAACACG GCGCATTGCC ATGTTGATCC CGGCTCCAAT 601 GACTATTTGC GCCGCATCGC CAGTTTTGCC AGTCAGCTCC ACAAGAATGC CTGCCAGCTT 661 GCGATCCTGC AGATAGAGGT CATTAGGCCA TTTGACACGA ACTTTATCTG CTCCCAGCTT 721 GCGTAATACT TCCGCCATCA CGATACCGAT AACCAGACTT AAACCAATCG CCGCCGCCGG 781 GCCTTGTTCC AGACGCCAGA ACATCGACAA ATATAAGTTT GCGCCAAAAG GCGAAAACCA 841 TTTCCGCCCC CGGCGACCAC GGCCAGCCTG CTGGTATTCT GCAACACAGG CATCGCCCGA 901 TTTAAGCTCT CCGATACGAT CAAGAAGGTA CTGATTCGTG GAGTCAATAA CTGGCAGCAC 961 GGCTACACTA CCGCCATCCA GCTGACCCAA TATCTGTTCA GCATTAAGTA ACTGGATGGG 1021 CTCAGGCAGG CTGTATCCTT TACCCGGAAC GGTAAAGACA TCAACGCCCC AGTCACGCAG 1081 TGTCTGAATG TGTTTATTAA TAGCCGCCCG GCTCATTCCC AGCGTTTCAC CCAACTGCTC 1141 GCCAGAGTGA AATTCACCGT TCGCTAACAG GGCAATCAAT TTCAGTGGCA CGGTGTTATC 1201 CTTCATGGGA GAGACTCCTT CTTAAAGTTA AGTTATCCAC TTCCAATATT GGATTGGAAG 1261 TACAGGTTCT CGGTACCTTC ATGCCACTCA ATTTTTTGCG CCTCAAAAAT ATCGTTCAGT 1321 CCAGAAGAAT GATGATGATG ATGGTGCATA TGTATATCTC CTTCTTAAAG TTAAACAAAA 1381 TTATTTCTAG AGGGGAATTG TTATCCGCTC ACAATTCCCC TATAGTGAGT CGTATTAATT 1441 TCGCGGGATC GAGATCGATC TCGATCCTCT ACGCCGGACG CATCGTGGCC GGCATCACCG 1501 GCGCCACAGG TGCGGTTGCT GGCGCCTATA TCGCCGACAT CACCGATGGG GAAGATCGGG 1561 CTCGCCACTT CGGGCTCATG AGCGCTTGTT TCGGCGTGGG TATGGTGGCA GGCCCCGTGG 1621 CCGGGGGACT GTTGGGCGCC ATCTCCTTGC ATGCACCATT CCTTGCGGCG GCGGTGCTCA 1681 ACGGCCTCAA CCTACTACTG GGCTGCTTCC TAATGCAGGA GTCGCATAAG GGAGAGCGTC 1741 GAGATCCCGG ACACCATCGA ATGGCGCAAA ACCTTTCGCG GTATGGCATG ATAGCGCCCG 1801 GAAGAGAGTC AATTCAGGGT GGTGAATGTG AAACCAGTAA CGTTATACGA TGTCGCAGAG 1861 TATGCCGGTG TCTCTTATCA GACCGTTTCC CGCGTGGTGA ACCAGGCCAG CCACGTTTCT 1921 GCGAAAACGC GGGAAAAAGT GGAAGCGGCG ATGGCGGAGC TGAATTACAT TCCCAACCGC 1981 GTGGCACAAC AACTGGCGGG CAAACAGTCG TTGCTGATTG GCGTTGCCAC CTCCAGTCTG 2041 GCCCTGCACG CGCCGTCGCA AATTGTCGCG GCGATTAAAT CTCGCGCCGA TCAACTGGGT 2101 GCCAGCGTGG TGGTGTCGAT GGTAGAACGA AGCGGCGTCG AAGCCTGTAA AGCGGCGGTG 2161 CACAATCTTC TCGCGCAACG CGTCAGTGGG CTGATCATTA ACTATCCGCT GGATGACCAG 2221 GATGCCATTG CTGTGGAAGC TGCCTGCACT AATGTTCCGG CGTTATTTCT TGATGTCTCT 2281 GACCAGACAC CCATCAACAG TATTATTTTC TCCCATGAAG ACGGTACGCG ACTGGGCGTG 2341 GAGCATCTGG TCGCATTGGG TCACCAGCAA ATCGCGCTGT TAGCGGGCCC ATTAAGTTCT 2401 GTCTCGGCGC GTCTGCGTCT GGCTGGCTGG CATAAATATC TCACTCGCAA TCAAATTCAG 2461 CCGATAGCGG AACGGGAAGG CGACTGGAGT GCCATGTCCG GTTTTCAACA AACCATGCAA 2521 ATGCTGAATG AGGGCATCGT TCCCACTGCG ATGCTGGTTG CCAACGATCA GATGGCGCTG 2581 GGCGCAATGC GCGCCATTAC CGAGTCCGGG CTGCGCGTTG GTGCGGATAT CTCGGTAGTG 2641 GGATACGACG ATACCGAAGA CAGCTCATGT TATATCCCGC CGTTAACCAC CATCAAACAG 2701 GATTTTCGCC TGCTGGGGCA AACCAGCGTG GACCGCTTGC TGCAACTCTC TCAGGGCCAG 2761 GCGGTGAAGG GCAATCAGCT GTTGCCCGTC TCACTGGTGA AAAGAAAAAC CACCCTGGCG 2821 CCCAATACGC AAACCGCCTC TCCCCGCGCG TTGGCCGATT CATTAATGCA GCTGGCACGA 2881 CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGCAACGCA ATTAATGTAA GTTAGCTCAC 2941 TCATTAGGCA CCGGGATCTC GACCGATGCC CTTGAGAGCC TTCAACCCAG TCAGCTCCTT 3001 CCGGTGGGCG CGGGGCATGA CTATCGTCGC CGCACTTATG ACTGTCTTCT TTATCATGCA 3061 ACTCGTAGGA CAGGTGCCGG CAGCGCTCTG GGTCATTTTC GGCGAGGACC GCTTTCGCTG 3121 GAGCGCGACG ATGATCGGCC TGTCGCTTGC GGTATTCGGA ATCTTGCACG CCCTCGCTCA 3181 AGCCTTCGTC ACTGGTCCCG CCACCAAACG TTTCGGCGAG AAGCAGGCCA TTATCGCCGG 3241 CATGGCGGCC CCACGGGTGC GCATGATCGT GCTCCTGTCG TTGAGGACCC GGCTAGGCTG 3301 GCGGGGTTGC CTTACTGGTT AGCAGAATGA ATCACCGATA CGCGAGCGAA CGTGAAGCGA 3361 CTGCTGCTGC AAAACGTCTG CGACCTGAGC AACAACATGA ATGGTCTTCG GTTTCCGTGT 3421 TTCGTAAAGT CTGGAAACGC GGAAGTCAGC GCCCTGCACC ATTATGTTCC GGATCTGCAT 3481 CGCAGGATGC TGCTGGCTAC CCTGTGGAAC ACCTACATCT GTATTAACGA AGCGCTGGCA 3541 TTGACCCTGA GTGATTTTTC TCTGGTCCCG CCGCATCCAT ACCGCCAGTT GTTTACCCTC 3601 ACAACGTTCC AGTAACCGGG CATGTTCATC ATCAGTAACC CGTATCGTGA GCATCCTCTC 3661 TCGTTTCATC GGTATCATTA CCCCCATGAA CAGAAATCCC CCTTACACGG AGGCATCAGT 3721 GACCAAACAG GAAAAAACCG CCCTTAACAT GGCCCGCTTT ATCAGAAGCC AGACATTAAC 3781 GCTTCTGGAG AAACTCAACG AGCTGGACGC GGATGAACAG GCAGACATCT GTGAATCGCT 3841 TCACGACCAC GCTGATGAGC TTTACCGCAG CTGCCTCGCG CGTTTCGGTG ATGACGGTGA 3901 AAACCTCTGA CACATGCAGC TCCCGGAGAC GGTCACAGCT TGTCTGTAAG CGGATGCCGG 3961 GAGCAGACAA GCCCGTCAGG GCGCGTCAGC GGGTGTTGGC GGGTGTCGGG GCGCAGCCAT 4021 GACCCAGTCA CGTAGCGATA GCGGAGTGTA TACTGGCTTA ACTATGCGGC ATCAGAGCAG 4081 ATTGTACTGA GAGTGCACCA TATATGCGGT GTGAAATACC GCACAGATGC GTAAGGAGAA 4141 AATACCGCAT CAGGCGCTCT TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC 4201 GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG 4261 GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA 4321 AGGCCGCGTT GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC 4381 GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC 4441 CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG 4501 CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT 4561 CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC 4621 GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC 4681 CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG 4741 AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG 4801 CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA 4861 CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG 4921 GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT 4981 CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA 5041 ATTAAAAATG AAGTTTTAAA TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT 5101 ACCAATGCTT AATCAGTGAG GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG 5161 TTGCCTGACT CCCCGTCGTG TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA 5221 GTGCTGCAAT GATACCGCGA GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC 5281 AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT 5341 CTATTAATTG TTGCCGGGAA GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG 5401 TTGTTGCCAT TGCTGCAGGC ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA 5461 GCTCCGGTTC CCAACGATCA AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG 5521 TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA 5581 TGGTTATGGC AGCACTGCAT AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG 5641 TGACTGGTGA GTACTCAACC AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT 5701 CTTGCCCGGC GTCAATACGG GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA 5761 TCATTGGAAA ACGTTCTTCG GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA 5821 GTTCGATGTA ACCCACTCGT GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG 5881 TTTCTGGGTG AGCAAAAACA GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC 5941 GGAAATGTTG AATACTCATA CTCTTCCTTT TTCAAATTAT TGAAGCATTT ATCAGGGTTA 6001 TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC 6061 GCGCACATTT CCCCGAAAAG TGCCACCTAA ATTGTAAGCG TTAATGTGAA CCATCACCCT 6121 AATCAAGTTT TTTGGGGTCG AGGTGCCGTA AAGCACTAAA TCGGAACCCT AAAGGGAGCC 6181 CCCGATTTAG AGCTTGACGG GGAAAGCCGG CGAACGTGGC GAGAAAGGAA GGGAAGAAAG 6241 CGAAAGGAGC GGGCGCTAGG GCGCTGGCAA GTGTAGCGGT CACGCTGCGC GTAACCACCA 6301 CACCCGCCGC GCTTAATGCG CCGCTACAGG GCGCGTCCCA TTCGCCA //