LOCUS V018713 8720 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018713 VERSION V018713 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..3400 /label="TMC2(NM_080751)" /note="TMC2(NM_080751)" /gene="TMC2" CDS 3407..3472 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 3496..4084 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 4150..4198 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4400..4828 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4842..5171 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5238..6032 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 6208..6329 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6378..6394) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6402..6418 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6426..6456) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6471..6492 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6780..7368) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7539..8399) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8400..8504) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatgagcca ccaggtaaag ggcctgaaag aggaagcacg 721 aggcggagtg aaagggcggg tgaagagcgg ctctccacac acaggtgaca ggctgggaag 781 gagatcctca agcaagcggg ctctcaaagc cgaggggacc ccaggcaggc gcggagctca 841 gcgaagccag aaggagcgcg ccgggggcag cccaagcccg gggtctcccc ggaggaagca 901 aacagggcgc aggagacaca gagaagagct gggggagcag gagcggggcg aggcagagag 961 gacctgcgag ggcaggagaa agcgcgacga gagggcctcc ttccaggagc ggacagcagc 1021 cccaaagagg gaaaaggaga ttccgaggag ggaggagaag tcgaagcggc agaagaaacc 1081 caggtcatcc tccttggcct ccagtgcctc tggtggggag tccctgtccg aggaggaact 1141 ggcccagatc ctggagcagg tggaagaaaa aaagaagctc attgccacca tgcggagcaa 1201 gccctggccc atggcgaaga agctgacaga gctcagggag gcccaggaat ttgtggagaa 1261 gtatgaaggt gccttgggaa aggggaaagg caagcaacta tatgcctaca agatgctgat 1321 ggccaagaaa tgggtcaaat ttaagagaga ctttgataat ttcaagactc aatgtatccc 1381 ctgggaaatg aagatcaagg acattgaaag tcactttggt tcttcagtgg catcgtattt 1441 catctttctc cgatggatgt atggagttaa ccttgtcctt tttggcttaa tatttggtct 1501 agtcataatc ccagaggtac tgatgggcat gccctatggg agtattccca gaaagacagt 1561 gcctcgggct gaggaagaaa aggccatgga tttttctgtc ctttgggatt ttgagggcta 1621 tatcaagtac tctgcactct tctatggcta ctacaacaac cagaggacca tcgggtggct 1681 gaggtaccgg ctgcctatgg cttactttat ggtgggggtc agcgtgttcg gctacagcct 1741 gattattgtc attcgatcga tggccagcaa tacccaagga agcacaggcg aaggggagag 1801 tgacaacttc acattcagct tcaagatgtt caccagctgg gactacctga tcgggaattc 1861 agagacagct gataacaaat atgcatccat caccaccagc ttcaaggaat caatagtgga 1921 tgaacaagag agtaacaaag aagaaaatat ccatctgaca agatttcttc gtgtcctggc 1981 caactttctc atcatctgct gtttgtgtgg aagtgggtac ctcatttact ttgtggttaa 2041 gcgatctcag caattctcca aaatgcagaa tgtcagctgg tatgaaagga atgaggtaga 2101 gatcgtgatg tccctgcttg gaatgttttg tccccctctg tttgaaacca tcgctgccct 2161 ggagaattac cacccacgca ctggactgaa gtggcagctg ggacgcatct ttgcactctt 2221 cctggggaac ctctacacat ttctcttggc cctgatggat gacgtccacc tcaagcttgc 2281 taatgaagag acaataaaga acatcactca ctggactctg tttaactatt acaactcttc 2341 tggttggaac gagagtgtcc cccgaccacc cctgcaccct gcagatgtgc cccggggttc 2401 ttgctgggag acagctgtgg gcattgaatt catgaggctg acggtgtctg acatgctggt 2461 aacgtacatc accatcctgc tgggggactt cctacgggct tgttttgtgc ggttcatgaa 2521 ctactgctgg tgctgggact tggaggctgg atttccttca tatgctgagt ttgatattag 2581 tggaaatgtg ctgggtttga tcttcaacca aggaatgatc tggatgggct ccttctatgc 2641 tccaggcctg gtgggcatta atgtgctgcg cctgctgacc tccatgtact tccagtgctg 2701 ggcggtgatg agcagcaacg taccccatga acgcgtgttc aaagcctccc gatccaacaa 2761 cttctacatg ggcctcctgc tgctggtgct cttcctcagc ctcctgccgg tggcctacac 2821 catcatgtcc ctcccaccct cctttgactg cgggccgttc agtgggaaaa acagaatgta 2881 cgatgtcctc caagagacca ttgaaaacga tttcccaacc ttcctgggca agatctttgc 2941 tttcctcgcc aatccaggcc tgatcatccc agccatcctg ctgatgttct tggccattta 3001 ctacctgaac tcagtttcca aaagcctttc ccgagctaat gcccagctga ggaagaaaat 3061 ccaagtgctc cgtgaagttg agaagagtca caaatctgta aaaggcaaag ccacagccag 3121 agattcagag gacacaccta aaagcagctc caaaaatgcc acccagctcc aactcaccaa 3181 ggaagagacc actcctccct ctgccagcca aagccaggcc atggacaaga aggcgcaggg 3241 ccctgggacc tccaattctg ccagcaggac cacactgcct gcctctggac accttcctat 3301 atctcggccc cctggaatcg gaccagattc tggccacgcc ccatctcaga ctcatccgtg 3361 gaggtcagcc tctggaaaga gtgctcagag acctccccac ctcgaggact acaaagacca 3421 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg 3481 gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 3541 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 3601 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 3661 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 3721 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 3781 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 3841 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 3901 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 3961 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 4021 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 4081 ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 4141 gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat 4201 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 4261 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 4321 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg 4381 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 4441 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 4501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc 4561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 4621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 4681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 4741 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag 4801 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4921 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 5041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 5101 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 5161 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag 5221 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 5281 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 5341 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 5401 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 5461 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 5521 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 5581 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 5641 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 5701 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 5761 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 5821 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 5881 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 5941 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 6001 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc 6061 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 6121 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 6181 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6301 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 6361 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 6421 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 6481 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 6541 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 6601 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 6661 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 6721 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 6781 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 6841 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 6901 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 6961 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 7021 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 7081 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 7141 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 7201 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 7261 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 7321 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 7381 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 7441 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 7501 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 7561 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 7621 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 7681 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 7741 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 7801 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 7861 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 7921 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 7981 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 8041 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 8101 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 8161 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 8221 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 8281 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 8341 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 8401 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 8461 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 8521 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 8581 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 8641 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 8701 gcatgaagaa tctgcttagg //