LOCUS pETDuetMOD3 5448 bp ds-DNA circular 20-MAY-2013 DEFINITION pETDUET-1 SOURCE ORGANISM COMMENT Dual gene/MCS expression vector with pBR322 (pET) origin with NcoI/BamHI fragment in MCS1 containing 6x His tag and 3c cut site COMMENT engineered by NPG 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(3560..3565) /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(3536..3542) /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 2065..2512 /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 4255..4255 /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(4686..4877) /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 1980..2027 /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(2634..3494) /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 1884..1928 /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 1753..1773 /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 1732..1748 /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 1804..1809 /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 1707..1726 /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(1707..1726) /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(1966..1984) /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 ATGGGTTCTT CTCACCATCA CCATCACCAT GGCAGCAGCC TGGAAGTTCT GTTCCAGGGG 1621 CCCGGATCCg aattcgagct cggcgcgcct gcaggtcgac aagcttgcgg ccgcataatg 1681 cttaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa ttaatacgac 1741 tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt agttaagtat 1801 aagaaggaga tatacatatg gcagatctca attggatatc ggccggccac gcgatcgctg 1861 acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca 1921 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac 1981 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa 2041 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 2101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 2161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2461 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc 2521 tggcggcacg atggcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 2581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 2641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 2701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 2761 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 2821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 2881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 2941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 3001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 3061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 3121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 3181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 3241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 3301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 3361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 3421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 3481 tgttgaatac tcatactctt cctttttcaa tcatgattga agcatttatc agggttattg 3541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gtcatgacca 3601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 3661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 3721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 3781 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 3841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 3901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 3961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 4021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 4081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 4141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 4201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 4261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 4321 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 4381 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 4441 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg 4501 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg 4561 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga 4621 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 4681 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca 4741 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg 4801 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt 4861 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg 4921 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg 4981 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa 5041 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc 5101 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt 5161 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac 5221 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca 5281 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgctagtc 5341 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga 5401 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgc //