COMMENT pMCSG51 5346 bp pMCSG50 truncated by digestion with PshAI and Tth111I followed by treatment with Klenow and dNTPs. This removes the rop repressor and flanking sequences. It was assumed that the PshAI site remained blunt and that the Tth111I site had the single-base overhang filled in by Klenow. 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(4098..4958) /note="bla cds" rep_origin complement(3339..3927) /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 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 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 CTACAGTCAC GTAGCGATAG CGGAGTGTAT ACTGGCTTAA 3061 CTATGCGGCA TCAGAGCAGA TTGTACTGAG AGTGCACCAT ATATGCGGTG TGAAATACCG 3121 CACAGATGCG TAAGGAGAAA ATACCGCATC AGGCGCTCTT CCGCTTCCTC GCTCACTGAC 3181 TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA 3241 CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA 3301 AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT 3361 GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA 3421 AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG 3481 CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCATAGCTCA 3541 CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA 3601 CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG 3661 GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG 3721 TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG 3781 ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC 3841 TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG 3901 ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC 3961 GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC 4021 TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG 4081 TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT 4141 CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG 4201 GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA 4261 GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT 4321 TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA 4381 GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTGCAGGCA TCGTGGTGTC ACGCTCGTCG 4441 TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC 4501 ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG 4561 GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA 4621 TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT 4681 ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC GCCACATAGC 4741 AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC 4801 TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA 4861 TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA 4921 AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAAATTATT 4981 GAAGCATTTA TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA 5041 ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTAAA TTGTAAGCGT 5101 TAATGTGAAC CATCACCCTA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT 5161 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG 5221 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC 5281 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTCCCAT 5341 TCGCCA //