LOCUS pETDuetMOD2 5478 bp ds-DNA circular 20-MAY-2013 DEFINITION pETDUETMOD2 SOURCE ORGANISM COMMENT pET-Duet1 with a NdeI-BglII fragment cloned into MCS2; Based on MCS from pBG100 COMMENT SM Shell COMMENT ApEinfo:methylated:0 FEATURES Location/Qualifiers RBS 1548..1553 /vntifkey="32" /label=RBS-1 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 -35_signal complement(3590..3595) /vntifkey="48" /label=-35\Ap /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 -10_signal complement(3566..3572) /vntifkey="47" /label=-10\Ap /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 rep_origin 2095..2542 /vntifkey="33" /label=f1\ori /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 rep_origin 4285..4285 /vntifkey="33" /label=ColE1,\pBR322\ori /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 CDS complement(4716..4907) /vntifkey="4" /label=ROP /ApEinfo_fwdcolor=#0000ff /ApEinfo_revcolor=#ffff00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 protein_bind 1495..1515 /vntifkey="31" /label=lacO /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 terminator 2010..2057 /vntifkey="43" /label=T7\terminator /ApEinfo_fwdcolor=#ff1e9e /ApEinfo_revcolor=#ff1e9e /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 CDS complement(2664..3524) /vntifkey="4" /note="B-lactamase gene" /label=Ap /ApEinfo_fwdcolor=#0000ff /ApEinfo_revcolor=#ffff00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_binding 1573..1590 /vntifkey="20" /label=His-Tag /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 protein_bind 1914..1958 /vntifkey="31" /label=S-Tag /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 protein_bind 1725..1745 /vntifkey="31" /label=lacO(1) /ApEinfo_label=lacO /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 promoter 1704..1720 /vntifkey="30" /label=T7\promoter /ApEinfo_fwdcolor=#007cff /ApEinfo_revcolor=#007c00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 RBS 1776..1781 /vntifkey="32" /label=RBS /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer 1679..1698 /vntifkey="27" /label=DuetUP2\Primer\#71180-3 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer complement(1679..1698) /vntifkey="27" /label=DuetDOWN1\Primer\#71179-3 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer complement(1996..2014) /vntifkey="27" /label=T7\Terminator\Primer\#69337-3 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 CDS complement(1..1083) /vntifkey="4" /note="lac repressor gene" /label=lacI /ApEinfo_fwdcolor=#0000ff /ApEinfo_revcolor=#ffff00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 promoter 1474..1490 /vntifkey="30" /label=T7\promoter(1) /ApEinfo_label=T7\promoter /ApEinfo_fwdcolor=#007cff /ApEinfo_revcolor=#007c00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer 1428..1443 /vntifkey="27" /label=pET\Upstream\Primer\#69214-3 /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 ORIGIN 1 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 61 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga 121 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc 181 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata 241 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag 301 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat 361 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc 421 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg 481 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat 541 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga 601 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt 661 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag 721 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct 781 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc 841 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa 901 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat 961 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg 1021 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt 1081 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt 1141 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta 1201 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat 1261 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga 1321 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga 1381 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt 1441 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt 1501 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc 1561 atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag ctcggcgcgc 1621 ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt 1681 gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt gagcggataa 1741 caattcccca tcttagtata ttagttaagt ataagaagga gatataCATA TGGGTTCTTC 1801 TCACCATCAC CATCACCATG GCAGCAGCCT GGAAGTTCTG TTCCAGGGGC CCAGATCTca 1861 attggatatc ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa 1921 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa 1981 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg 2041 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc 2101 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2161 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2221 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2281 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2341 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2401 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2461 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2521 ttttaacaaa atattaacgt ttacaatttc tggcggcacg atggcatgag attatcaaaa 2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 2701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 2761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 2821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 2881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 2941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 3001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 3061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 3121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 3181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 3241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 3301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 3361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 3421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 3481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 3541 tcatgattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 3601 ttagaaaaat aaacaaatag gtcatgacca aaatccctta acgtgagttt tcgttccact 3661 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 3721 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 3781 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 3841 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 3901 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 3961 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 4021 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 4081 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 4141 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 4201 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 4261 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 4321 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 4381 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 4441 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 4501 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc 4561 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 4621 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 4681 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 4741 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg 4801 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt 4861 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg 4921 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg 4981 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg 5041 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt 5101 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac 5161 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag 5221 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc 5281 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc 5341 ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa ggagctgact 5401 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt 5461 acattaattg cgttgcgc //