LOCUS pETDuetMOD1 5439 bp ds-DNA circular 20-MAY-2013 DEFINITION pETDUETMOD1 SOURCE ORGANISM COMMENT pET-Duet1 with an NcoI-BamHI fragment cloned into MCS1; based on MCS from pSV281 with MGSSHHHHHHENLYFQGS (his/tev), BamHI clones, in frame COMMENT From SM Shell COMMENT 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(3551..3556) /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(3527..3533) /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 2056..2503 /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 4246..4246 /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(4677..4868) /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 1971..2018 /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(2625..3485) /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 protein_bind 1875..1919 /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 1744..1764 /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 1723..1739 /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 1795..1800 /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 1698..1717 /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(1698..1717) /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(1957..1975) /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 AGCCAGGAAA ACCTGTACTT CCAGGGATCC 1621 gaattcgagc tcggcgcgcc tgcaggtcga caagcttgcg gccgcataat gcttaagtcg 1681 aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag 1741 gggaattgtg agcggataac aattccccat cttagtatat tagttaagta taagaaggag 1801 atatacatat ggcagatctc aattggatat cggccggcca cgcgatcgct gacgtcggta 1861 ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt 1921 ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac 1981 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg 2041 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2101 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 2221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 2341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 2401 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 2461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt ctggcggcac 2521 gatggcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 2581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 2641 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 2701 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 2761 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 2821 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 2881 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 2941 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 3001 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 3061 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 3121 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 3181 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 3241 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 3301 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 3361 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 3421 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 3481 ctcatactct tcctttttca atcatgattg aagcatttat cagggttatt gtctcatgag 3541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggtcatgacc aaaatccctt 3601 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 3661 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 3721 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 3781 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca 3841 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 3901 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 3961 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4021 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4081 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4141 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4201 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 4261 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 4321 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 4381 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc 4441 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag 4501 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 4561 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 4621 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 4681 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg 4741 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc 4801 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt 4861 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg 4921 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa 4981 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag 5041 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat 5101 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt 5161 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag 5221 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa 5281 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgctagt catgccccgc 5341 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt 5401 gcctaatgag tgagctaact tacattaatt gcgttgcgc //