COMMENT pMCSG53 4808 bp base vector: pMCSG52 pMCSG52: argU tRNA gene inserted into the SgrAI site of pMCSG7 ileX tRNA gene inserted into the SphI site of pMCSG7 [argU: Garcia et al. 1986 Cell 45:453-459] [ileX: del Tito et al. 1995 J. Bacteriol. 177:7086-7091] pMCSG53: 1 kb of sequence (including the rop repressor) removed from pMCSG52 by digestion with PshAI and Tth111I, followed by treatment with Klenow, dNTPs and T4 DNA ligase FEATURES Location/Qualifiers tRNA complement(959..1110) /note="ileX gene" rep_origin complement(2801..3389) /note="pBR322 origin of replication" misc_feature complement(204..235) /note="LIC Site" Cleavage-site complement(223..243) /note="TEV Site" Region complement(268..285) /note="His Tag" promoter complement(360..376) /note="T7 Promoter" terminator complement(26..72) /note="T7 Terminator" CDS 1413..2372 /note="Lac I cds" CDS complement(3560..4420) /note="bla cds" tRNA complement(443..799) /note="argU tRNA" 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 ACTTCCAATA TTGGATTGGA AGTACAGGTT 241 CTCGGTACCC AGATCTACAC CAGAAGAATG ATGATGATGA TGGTGCATAT GTATATCTCC 301 TTCTTAAAGT TAAACAAAAT TATTTCTAGA GGGGAATTGT TATCCGCTCA CAATTCCCCT 361 ATAGTGAGTC GTATTAATTT CGCGGGATCG AGATCGATCT CGATCCTCTA CGCCGGACGC 421 ATCGTGGCCG GCATCACCGG TGGATTCAGT CAGGCGTCCC ATTATCAGTG CTTCAGGAAA 481 TGGGCGGATG GGAGTCCATA GAAATGGTTC GTAGGTATGC TCACCTTGCG CCTAATCATT 541 TGACAGAGCA TGCGAGGAAA ATAGACGACA TTTTTGGTGA TAATGTCCCA AATATGTCCC 601 ACTCTGGAAT TATGGAGGAT ATAAAGAAGG CGTAACTGAT TGAATTGTAA TGGCGCGCCC 661 TGCAGGATTC GAACCTGCGG CCCACGACTT AGAAGGTCGT TGCTCTATCC AACTGAGCTA 721 AGGGCGCGTT GATACCGCAA TGCGGTGTAA TCGCGTGAAT TATACGGTCA ACCCTTGCTG 781 AGTCAATGGC TTTTGATCTC GCCGGCGCCA CAGGTGCGGT TGCTGGCGCC TATATCGCCG 841 ACATCACCGA TGGGGAAGAT CGGGCTCGCC ACTTCGGGCT CATGAGCGCT TGTTTCGGCG 901 TGGGTATGGT GGCAGGCCCC GTGGCCGGGG GACTGTTGGG CGCCATCTCC TTGCATGCAA 961 AAAGGCTGAC GATTTCTCGT CAGCCTTTGC TATATCTGGT GGCCCCTGCT GGACTTGAAC 1021 CAGCGACCAA GCGATTATGA GTCGCCTGCT CTAACCACTG AGCTAAGGGG CCATGGTAGC 1081 GGATTATAAA GTAACTCCGT GTCGCAATCC GCATGCACCA TTCCTTGCGG CGGCGGTGCT 1141 CAACGGCCTC AACCTACTAC TGGGCTGCTT CCTAATGCAG GAGTCGCATA AGGGAGAGCG 1201 TCGAGATCCC GGACACCATC GAATGGCGCA AAACCTTTCG CGGTATGGCA TGATAGCGCC 1261 CGGAAGAGAG TCAATTCAGG GTGGTGAATG TGAAACCAGT AACGTTATAC GATGTCGCAG 1321 AGTATGCCGG TGTCTCTTAT CAGACCGTTT CCCGCGTGGT GAACCAGGCC AGCCACGTTT 1381 CTGCGAAAAC GCGGGAAAAA GTGGAAGCGG CGATGGCGGA GCTGAATTAC ATTCCCAACC 1441 GCGTGGCACA ACAACTGGCG GGCAAACAGT CGTTGCTGAT TGGCGTTGCC ACCTCCAGTC 1501 TGGCCCTGCA CGCGCCGTCG CAAATTGTCG CGGCGATTAA ATCTCGCGCC GATCAACTGG 1561 GTGCCAGCGT GGTGGTGTCG ATGGTAGAAC GAAGCGGCGT CGAAGCCTGT AAAGCGGCGG 1621 TGCACAATCT TCTCGCGCAA CGCGTCAGTG GGCTGATCAT TAACTATCCG CTGGATGACC 1681 AGGATGCCAT TGCTGTGGAA GCTGCCTGCA CTAATGTTCC GGCGTTATTT CTTGATGTCT 1741 CTGACCAGAC ACCCATCAAC AGTATTATTT TCTCCCATGA AGACGGTACG CGACTGGGCG 1801 TGGAGCATCT GGTCGCATTG GGTCACCAGC AAATCGCGCT GTTAGCGGGC CCATTAAGTT 1861 CTGTCTCGGC GCGTCTGCGT CTGGCTGGCT GGCATAAATA TCTCACTCGC AATCAAATTC 1921 AGCCGATAGC GGAACGGGAA GGCGACTGGA GTGCCATGTC CGGTTTTCAA CAAACCATGC 1981 AAATGCTGAA TGAGGGCATC GTTCCCACTG CGATGCTGGT TGCCAACGAT CAGATGGCGC 2041 TGGGCGCAAT GCGCGCCATT ACCGAGTCCG GGCTGCGCGT TGGTGCGGAT ATCTCGGTAG 2101 TGGGATACGA CGATACCGAA GACAGCTCAT GTTATATCCC GCCGTTAACC ACCATCAAAC 2161 AGGATTTTCG CCTGCTGGGG CAAACCAGCG TGGACCGCTT GCTGCAACTC TCTCAGGGCC 2221 AGGCGGTGAA GGGCAATCAG CTGTTGCCCG TCTCACTGGT GAAAAGAAAA ACCACCCTGG 2281 CGCCCAATAC GCAAACCGCC TCTCCCCGCG CGTTGGCCGA TTCATTAATG CAGCTGGCAC 2341 GACAGGTTTC CCGACTGGAA AGCGGGCAGT GAGCGCAACG CAATTAATGT AAGTTAGCTC 2401 ACTCATTAGG CACCGGGATC TCGACCGATG CCCTTGAGAG CCTTCAACCC AGTCAGCTCC 2461 TTCCGGTGGG CGCGGGGCAT GACTACAGTC ACGTAGCGAT AGCGGAGTGT ATACTGGCTT 2521 AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC ATATATGCGG TGTGAAATAC 2581 CGCACAGATG CGTAAGGAGA AAATACCGCA TCAGGCGCTC TTCCGCTTCC TCGCTCACTG 2641 ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC GAGCGGTATC AGCTCACTCA AAGGCGGTAA 2701 TACGGTTATC CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA AAAGGCCAGC 2761 AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG CTCCGCCCCC 2821 CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG ACAGGACTAT 2881 AAAGATACCA GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT CCGACCCTGC 2941 CGCTTACCGG ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT TCTCATAGCT 3001 CACGCTGTAG GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC TGTGTGCACG 3061 AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT GAGTCCAACC 3121 CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT AGCAGAGCGA 3181 GGTATGTAGG CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC TACACTAGAA 3241 GGACAGTATT TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA 3301 GCTCTTGATC CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC 3361 AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT ACGGGGTCTG 3421 ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAGATTA TCAAAAAGGA 3481 TCTTCACCTA GATCCTTTTA AATTAAAAAT GAAGTTTTAA ATCAATCTAA AGTATATATG 3541 AGTAAACTTG GTCTGACAGT TACCAATGCT TAATCAGTGA GGCACCTATC TCAGCGATCT 3601 GTCTATTTCG TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT ACGATACGGG 3661 AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGACCCACGC TCACCGGCTC 3721 CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA GCGCAGAAGT GGTCCTGCAA 3781 CTTTATCCGC CTCCATCCAG TCTATTAATT GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC 3841 CAGTTAATAG TTTGCGCAAC GTTGTTGCCA TTGCTGCAGG CATCGTGGTG TCACGCTCGT 3901 CGTTTGGTAT GGCTTCATTC AGCTCCGGTT CCCAACGATC AAGGCGAGTT ACATGATCCC 3961 CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC AGAAGTAAGT 4021 TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA TAATTCTCTT ACTGTCATGC 4081 CATCCGTAAG ATGCTTTTCT GTGACTGGTG AGTACTCAAC CAAGTCATTC TGAGAATAGT 4141 GTATGCGGCG ACCGAGTTGC TCTTGCCCGG CGTCAATACG GGATAATACC GCGCCACATA 4201 GCAGAACTTT AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA CTCTCAAGGA 4261 TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC TGATCTTCAG 4321 CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC AGGAAGGCAA AATGCCGCAA 4381 AAAAGGGAAT AAGGGCGACA CGGAAATGTT GAATACTCAT ACTCTTCCTT TTTCAAATTA 4441 TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT GTATTTAGAA 4501 AAATAAACAA ATAGGGGTTC CGCGCACATT TCCCCGAAAA GTGCCACCTA AATTGTAAGC 4561 GTTAATGTGA ACCATCACCC TAATCAAGTT TTTTGGGGTC GAGGTGCCGT AAAGCACTAA 4621 ATCGGAACCC TAAAGGGAGC CCCCGATTTA GAGCTTGACG GGGAAAGCCG GCGAACGTGG 4681 CGAGAAAGGA AGGGAAGAAA GCGAAAGGAG CGGGCGCTAG GGCGCTGGCA AGTGTAGCGG 4741 TCACGCTGCG CGTAACCACC ACACCCGCCG CGCTTAATGC GCCGCTACAG GGCGCGTCCC 4801 ATTCGCCA //