LOCUS Exported 7389 bp ds-DNA circular SYN 27-APR-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7389) AUTHORS Weatherall Institute TITLE Direct Submission JOURNAL Exported Nov 30, 2018 from SnapGene Viewer 4.2.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7389 /organism="recombinant plasmid" /mol_type="other DNA" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 911..935 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 919..949 /label=PCR8.1 misc_feature 950..955 /label=Shine dalgarno sequence misc_feature 956..961 /label=Spacer misc_feature 965..2815 /label=NS3 misc_feature 2816..2838 /label=PCR8.2 protein_bind complement(2829..2853) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2879..2920 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 3004..3052 /label=HSV TK poly(A) signal /note="herpesvirus thymidine kinase polyadenylation signal" rep_origin 3254..3682 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3696..4025 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3876..4011 /label=SV40 ori /note="SV40 origin of replication" CDS 4092..4886 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 5062..5183 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(5634..6222) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6393..7253) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7254..7358) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggctccgaa ttcgccctta ggaggacagc 961 tatgagtggt gctctatggg atgtgcctgc tcccaaggaa gtaaaaaagg gggagaccac 1021 agatggagtg tacagagtaa tgactcgtag actgctaggt tcaacacaag ttggagtggg 1081 agttatgcaa gagggggtct ttcacactat gtggcacgtc acaaaaggat ccgcgctgag 1141 aagcggtgaa gggagacttg atccatactg gggagatgtc aagcaggatc tggtgtcata 1201 ctgtggtcca tggaagctag atgccgcctg ggacgggcac agcgaggtgc agctcttggc 1261 cgtgcccccc ggagagagag cgaggaacat ccagactctg cccggaatat ttaagacaaa 1321 ggatggggac attggagcgg ttgcgctgga ttacccagca ggaacttcag gatctccaat 1381 cctagacaag tgtgggagag tgataggact ttatggcaat ggggtcgtga tcaaaaatgg 1441 gagttatgtt agtgccatca cccaagggag gagggaggaa gagactcctg ttgagtgctt 1501 cgagccttcg atgctgaaga agaagcagct aactgtctta gacttgcatc ctggagctgg 1561 gaaaaccagg agagttcttc ctgaaatagt ccgtgaagcc ataaaaacaa gactccgtac 1621 tgtgatctta gctccaacca gggttgtcgc tgctgaaatg gaggaagccc ttagagggct 1681 tccagtgcgt tatatgacaa cagcagtcaa tgtcacccac tctggaacag aaatcgtcga 1741 cttaatgtgc catgccacct tcacttcacg tctactacag ccaatcagag tccccaacta 1801 taatctgtat attatggatg aggcccactt cacagatccc tcaagtatag cagcaagagg 1861 atacatttca acaagggttg agatgggcga ggcggctgcc atcttcatga ccgccacgcc 1921 accaggaacc cgtgacgcat ttccggactc caactcacca attatggaca ccgaagtgga 1981 agtcccagag agagcctgga gctcaggctt tgattgggtg acggatcatt ctggaaaaac 2041 agtttggttt gttccaagcg tgaggaacgg caatgagatc gcagcttgtc tgacaaaggc 2101 tggaaaacgg gtcatacagc tcagcagaaa gacttttgag acagagttcc agaaaacaaa 2161 acatcaagag tgggactttg tcgtgacaac tgacatttca gagatgggcg ccaactttaa 2221 agctgaccgt gtcatagatt ccaggagatg cctaaagccg gtcatacttg atggcgagag 2281 agtcattttg gctggaccca tgcctgtcac acatgccagc gctgcccaga ggagggggcg 2341 cataggcagg aatcccaaca aacctggaga tgagtatctg tatggaggtg ggtgcgcaga 2401 gactgacgaa gaccatgcac actggcttga agcaagaatg ctccttgaca atatttacct 2461 ccaagatggc ctcatagcct cgctctatcg acctgaggcc gacaaagtag cagccattga 2521 gggagagttc aagcttagga cggagcaaag gaagaccttt gtggaactca tgaaaagagg 2581 agatcttcct gtttggctgg cctatcaggt tgcatctgcc ggaataacct acacagatag 2641 aagatggtgc tttgatggca cgaccaacaa caccataatg gaagacagtg tgccggcaga 2701 ggtgtggacc agacacggag agaaaagagt gctcaaaccg aggtggatgg acgccagagt 2761 ttgttcagat catgcggccc tgaagtcatt caaggagttt gccgctggga aaagaaaggg 2821 cgaattcgac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg 2881 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt 2941 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 3001 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 3061 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3121 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3181 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 3241 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3301 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3361 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 3421 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 3481 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3541 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 3601 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 3661 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 3721 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3841 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3961 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4021 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4261 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 4921 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 4981 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 5041 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 5101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 5161 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 5221 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 5461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5821 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6061 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7381 cctgacgtc //