LOCUS pRSFDuetMOD1 3857 bp ds-DNA circular 20-MAY-2013 DEFINITION pRSFDuet-1/Kan. SOURCE ORGANISM COMMENT Dual gene/MCS expression vector with RSF origin with NcoI/BamHI fragment in MCS1 contain 6x His tag and 3c cut site COMMENT originally engineered by S. Vaithiyalingam, re-engineered by NPG COMMENT ApEinfo:methylated:0 FEATURES Location/Qualifiers CDS complement(1999..2814) /vntifkey="4" /label=Kan /ApEinfo_fwdcolor=#0000ff /ApEinfo_revcolor=#ffff00 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 protein_bind 1237..1257 /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 RBS 1290..1295 /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 primer complement(1708..1726) /vntifkey="27" /label=T7\Terminator\primer /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 terminator 1722..1769 /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 protein_bind 1626..1670 /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 CDS 1560..1688 /vntifkey="4" /label=MCS-2\ORF /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(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 1474..1490 /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 1546..1551 /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 misc_RNA complement(2922..3620) /vntifkey="53" /label=RNA\II /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_RNA 3511..3617 /vntifkey="53" /label=RNA\I /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 rep_origin 2922..3671 /vntifkey="33" /label=RSF\ori\-\RSF1030\Homology /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 -10_signal 3499..3504 /vntifkey="47" /label=-10\RNA\I /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 -35_signal 3475..3480 /vntifkey="48" /label=-35\RNA\I /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 RBS complement(2820..2824) /vntifkey="32" /label=RBS(1) /ApEinfo_label=RBS /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 -35_signal complement(2879..2884) /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(2856..2862) /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 CDS complement(10..1092) /vntifkey="4" /note="Wildtype lacI 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 1216..1232 /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 ORIGIN 1 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 61 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt 121 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag 181 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg 241 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc 301 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc 361 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc 421 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag 481 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa 541 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc 601 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc 661 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt 721 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc 781 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga 841 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc 901 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag 961 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt 1021 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt 1081 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc 1141 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact 1201 cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc 1261 ctgtagaaat aattttgttt aactttaata aggagatata CCATGGGTTC TTCTCACCAT 1321 CACCATCACC ATGGCAGCAG CCTGGAAGTT CTGTTCCAGG GGCCCGGATC Cgaattcgag 1381 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag 1441 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt 1501 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata 1561 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt 1621 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg 1681 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg 1741 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg 1801 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt 1861 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt 1921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 1981 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt 2041 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa 2101 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg 2161 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa 2221 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca 2281 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc 2341 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca 2401 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt 2461 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt 2521 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat 2581 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc 2641 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt 2701 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat 2761 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt 2821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 2881 tgaatgtatt tagaaaaata aacaaatagg catgcagcgc tcttccgctt cctcgctcac 2941 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg tcagctcact caaaagcggt 3001 aatacggtta tccacagaat caggggataa agccggaaag aacatgtgag caaaaagcaa 3061 agcaccggaa gaagccaacg ccgcaggcgt ttttccatag gctccgcccc cctgacgagc 3121 atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc gacaggacta taaagatacc 3181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt 3301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3421 acgacttatc gccactggca gcagccattg gtaactgatt tagaggactt tgtcttgaag 3481 ttatgcacct gttaaggcta aactgaaaga acagattttg gtgagtgcgg tcctccaacc 3541 cacttacctt ggttcaaaga gttggtagct cagcgaacct tgagaaaacc accgttggta 3601 gcggtggttt ttctttattt atgagatgat gaatcaatcg gtctatcaag tcaacgaaca 3661 gctattccgt tactctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc 3721 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg 3781 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga 3841 gctaacttac attaatt //