COMMENT pMCSG62 5869 bp base vector: pMCSG53 LIC region, BirA gene and a portion of the beta-lactamase gene from pMCSG51 removed by digestion with BglI and XbaI then ligated into pMCSG53 digested with the same enzymes. Product is biotinylation vector with truncated ori and the argU & ileX tRNA genes FEATURES Location/Qualifiers tRNA complement(2020..2171) /note="ileX gene" rep_origin complement(3862..4450) /note="pBR322 origin of replication" promoter complement(1421..1437) /note="T7 Promoter" CDS 2474..3433 /note="Lac I cds" CDS complement(4621..5481) /note="bla cds" tRNA complement(1504..1860) /note="argU tRNA" misc_feature complement(1232..1263) /note="MCSG7 LIC site" Cleavage-site complement(1251..1271) /note="TEV Site" Region complement(1329..1346) /note="His Tag" terminator complement(26..72) /note="T7 Terminator" 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 complement(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 GTGGATTCAG TCAGGCGTCC CATTATCAGT GCTTCAGGAA ATGGGCGGAT GGGAGTCCAT 1561 AGAAATGGTT CGTAGGTATG CTCACCTTGC GCCTAATCAT TTGACAGAGC ATGCGAGGAA 1621 AATAGACGAC ATTTTTGGTG ATAATGTCCC AAATATGTCC CACTCTGGAA TTATGGAGGA 1681 TATAAAGAAG GCGTAACTGA TTGAATTGTA ATGGCGCGCC CTGCAGGATT CGAACCTGCG 1741 GCCCACGACT TAGAAGGTCG TTGCTCTATC CAACTGAGCT AAGGGCGCGT TGATACCGCA 1801 ATGCGGTGTA ATCGCGTGAA TTATACGGTC AACCCTTGCT GAGTCAATGG CTTTTGATCT 1861 CGCCGGCGCC ACAGGTGCGG TTGCTGGCGC CTATATCGCC GACATCACCG ATGGGGAAGA 1921 TCGGGCTCGC CACTTCGGGC TCATGAGCGC TTGTTTCGGC GTGGGTATGG TGGCAGGCCC 1981 CGTGGCCGGG GGACTGTTGG GCGCCATCTC CTTGCATGCA AAAAGGCTGA CGATTTCTCG 2041 TCAGCCTTTG CTATATCTGG TGGCCCCTGC TGGACTTGAA CCAGCGACCA AGCGATTATG 2101 AGTCGCCTGC TCTAACCACT GAGCTAAGGG GCCATGGTAG CGGATTATAA AGTAACTCCG 2161 TGTCGCAATC CGCATGCACC ATTCCTTGCG GCGGCGGTGC TCAACGGCCT CAACCTACTA 2221 CTGGGCTGCT TCCTAATGCA GGAGTCGCAT AAGGGAGAGC GTCGAGATCC CGGACACCAT 2281 CGAATGGCGC AAAACCTTTC GCGGTATGGC ATGATAGCGC CCGGAAGAGA GTCAATTCAG 2341 GGTGGTGAAT GTGAAACCAG TAACGTTATA CGATGTCGCA GAGTATGCCG GTGTCTCTTA 2401 TCAGACCGTT TCCCGCGTGG TGAACCAGGC CAGCCACGTT TCTGCGAAAA CGCGGGAAAA 2461 AGTGGAAGCG GCGATGGCGG AGCTGAATTA CATTCCCAAC CGCGTGGCAC AACAACTGGC 2521 GGGCAAACAG TCGTTGCTGA TTGGCGTTGC CACCTCCAGT CTGGCCCTGC ACGCGCCGTC 2581 GCAAATTGTC GCGGCGATTA AATCTCGCGC CGATCAACTG GGTGCCAGCG TGGTGGTGTC 2641 GATGGTAGAA CGAAGCGGCG TCGAAGCCTG TAAAGCGGCG GTGCACAATC TTCTCGCGCA 2701 ACGCGTCAGT GGGCTGATCA TTAACTATCC GCTGGATGAC CAGGATGCCA TTGCTGTGGA 2761 AGCTGCCTGC ACTAATGTTC CGGCGTTATT TCTTGATGTC TCTGACCAGA CACCCATCAA 2821 CAGTATTATT TTCTCCCATG AAGACGGTAC GCGACTGGGC GTGGAGCATC TGGTCGCATT 2881 GGGTCACCAG CAAATCGCGC TGTTAGCGGG CCCATTAAGT TCTGTCTCGG CGCGTCTGCG 2941 TCTGGCTGGC TGGCATAAAT ATCTCACTCG CAATCAAATT CAGCCGATAG CGGAACGGGA 3001 AGGCGACTGG AGTGCCATGT CCGGTTTTCA ACAAACCATG CAAATGCTGA ATGAGGGCAT 3061 CGTTCCCACT GCGATGCTGG TTGCCAACGA TCAGATGGCG CTGGGCGCAA TGCGCGCCAT 3121 TACCGAGTCC GGGCTGCGCG TTGGTGCGGA TATCTCGGTA GTGGGATACG ACGATACCGA 3181 AGACAGCTCA TGTTATATCC CGCCGTTAAC CACCATCAAA CAGGATTTTC GCCTGCTGGG 3241 GCAAACCAGC GTGGACCGCT TGCTGCAACT CTCTCAGGGC CAGGCGGTGA AGGGCAATCA 3301 GCTGTTGCCC GTCTCACTGG TGAAAAGAAA AACCACCCTG GCGCCCAATA CGCAAACCGC 3361 CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA CGACAGGTTT CCCGACTGGA 3421 AAGCGGGCAG TGAGCGCAAC GCAATTAATG TAAGTTAGCT CACTCATTAG GCACCGGGAT 3481 CTCGACCGAT GCCCTTGAGA GCCTTCAACC CAGTCAGCTC CTTCCGGTGG GCGCGGGGCA 3541 TGACTACAGT CACGTAGCGA TAGCGGAGTG TATACTGGCT TAACTATGCG GCATCAGAGC 3601 AGATTGTACT GAGAGTGCAC CATATATGCG GTGTGAAATA CCGCACAGAT GCGTAAGGAG 3661 AAAATACCGC ATCAGGCGCT CTTCCGCTTC CTCGCTCACT GACTCGCTGC GCTCGGTCGT 3721 TCGGCTGCGG CGAGCGGTAT CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC 3781 AGGGGATAAC GCAGGAAAGA ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA 3841 AAAGGCCGCG TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA 3901 TCGACGCTCA AGTCAGAGGT GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC 3961 CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC 4021 CGCCTTTCTC CCTTCGGGAA GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG 4081 TTCGGTGTAG GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA 4141 CCGCTGCGCC TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC 4201 GCCACTGGCA GCAGCCACTG GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC 4261 AGAGTTCTTG AAGTGGTGGC CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG 4321 CGCTCTGCTG AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA 4381 AACCACCGCT GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA 4441 AGGATCTCAA GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA 4501 CTCACGTTAA GGGATTTTGG TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT 4561 AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG 4621 TTACCAATGC TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT 4681 AGTTGCCTGA CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC 4741 CAGTGCTGCA ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA 4801 CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA 4861 GTCTATTAAT TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA 4921 CGTTGTTGCC ATTGCTGCAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT 4981 CAGCTCCGGT TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC 5041 GGTTAGCTCC TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT 5101 CATGGTTATG GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC 5161 TGTGACTGGT GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG 5221 CTCTTGCCCG GCGTCAATAC GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT 5281 CATCATTGGA AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC 5341 CAGTTCGATG TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG 5401 CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC 5461 ACGGAAATGT TGAATACTCA TACTCTTCCT TTTTCAAATT ATTGAAGCAT TTATCAGGGT 5521 TATTGTCTCA TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA AATAGGGGTT 5581 CCGCGCACAT TTCCCCGAAA AGTGCCACCT AAATTGTAAG CGTTAATGTG AACCATCACC 5641 CTAATCAAGT TTTTTGGGGT CGAGGTGCCG TAAAGCACTA AATCGGAACC CTAAAGGGAG 5701 CCCCCGATTT AGAGCTTGAC GGGGAAAGCC GGCGAACGTG GCGAGAAAGG AAGGGAAGAA 5761 AGCGAAAGGA GCGGGCGCTA GGGCGCTGGC AAGTGTAGCG GTCACGCTGC GCGTAACCAC 5821 CACACCCGCC GCGCTTAATG CGCCGCTACA GGGCGCGTCC CATTCGCCA //