LOCUS V029999 6393 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029999 VERSION V029999 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1044..2540 /label="Chrna3(NM_145129)" /note="Chrna3(NM_145129)" /gene="Chrna3" CDS 2559..2588 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2607..2630 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2650..2666) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2677..3299 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3448..4036) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4365..4412 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4644..5438) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" promoter complement(5473..5830) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5832..5936) /gene="bla" /label="AmpR promoter" rep_origin 5963..6391 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat gcggtcttca gacatgggtg ttgtgctgcc cccgccgccg ctgtccatgc 1081 tgatgctggt gctgatgctg ctgccagtgg ccagcgcctc ggaagctgag caccgcctgt 1141 tccagtacct gtttgaagat tacaacgaga tcatccggcc ggtggctaac gtgtcccatc 1201 ctgtcatcat ccagtttgag gtgtctatgt ctcagctggt gaaggtggat gaagtaaacc 1261 agatcatgga aaccaacctg tggctgaagc aaatctggaa tgactacaag ctgaaatgga 1321 aaccctctga ctaccaaggg gtggagttca tgcgagtccc tgcagagaag atctggaaac 1381 cagacatcgt gctttacaac aacgccgatg gggatttcca agtggatgac aaaaccaaag 1441 ctctactcaa gtacacagga gaagtgactt ggatccctcc ggccatcttt aagagctcat 1501 gcaaaatcga tgtgacctac ttcccgtttg actaccaaaa ctgcaccatg aagttcggct 1561 cctggtccta cgacaaggca aagatcgacc tggtcctcat tggctcctcc atgaacctca 1621 aggactattg ggaaagtggc gagtgggcca tcattaaagc cccgggctac aaacatgaaa 1681 tcaagtacaa ctgctgtgag gagatctacc aagacatcac gtactcgcta tacattcgcc 1741 gcctgccgct gttctacacc atcaacctca tcattccgtg cctgctcatc tccttcctca 1801 ctgtgctcgt cttctacctg ccctccgact gtggggagaa ggtgacgctc tgcatctccg 1861 tgctcctctc cctgacggtc tttctcctcg tgatcaccga gaccatccct tccacctcac 1921 tggtcatccc cttgatcggg gagtacctcc tcttcactat gatttttgtc accttgtcca 1981 tcgtcatcac agtctttgtg ctcaacgtgc actacagaac tccgaccaca cacacgatgc 2041 ccacttgggt caaggctgtg tttttgaacc ttctccccag ggtcatgttt atgactaggc 2101 caaccagcac cgaggaagac gccccaaaga cgaggaactt ctacggtgct gagctctcaa 2161 acctgaactg cttcagccgt gcagactcca aaagctgcaa ggaaggctac ccctgccaag 2221 atgggacatg tggctattgc caccaccgta gggtaaaaat ctcaaatttc agtgccaacc 2281 tcacaagaag ctccagttct gagtctgttg atgctgtgtt gtccctgtct gctctgtcac 2341 cagaaatcaa agaagccatc caaagtgtga agtatattgc tgaaaatatg aaagcacaga 2401 atgtagccaa agagattcaa gatgattgga agtatgttgc catggtgatt gatcgcattt 2461 ttctctgggt tttcatcctg gtgtgcattt taggaactgc aggattattt ctgcaaccct 2521 tgatggccag agatgacaca acgcgtacgc ggccgctcga gcagaaactc atctcagaag 2581 aggatctggc agcaaatgat atcctggatt acaaggatga cgacgataag gtttaaacgg 2641 ccggccgcgg tcatagctgt ttcctgaaca gatcccgggt ggcatccctg tgacccctcc 2701 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 2761 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 2821 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat 2881 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg 2941 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc 3001 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg 3061 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta 3121 caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag 3181 gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt 3241 tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg 3301 ggtacgcggc cagcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 3361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 3421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 3481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 3541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 3601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 3661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 3781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 3841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 3901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 3961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 4021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 4081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 4141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aacctgaggc 4201 tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg 4261 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg 4321 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac 4381 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt 4441 ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc 4501 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc 4561 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt 4621 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg 4681 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg 4741 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg atccgccaca 4801 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc 4861 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc 4921 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg 4981 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg 5041 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat 5101 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat 5161 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc 5221 gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac 5281 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca 5341 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg 5401 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc 5461 tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa 5521 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg 5581 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg 5641 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc 5701 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc 5761 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gctggttctt 5821 tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 5881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 5941 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 6001 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 6061 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 6121 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 6181 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 6241 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 6301 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 6361 gagctgattt aacaaaaatt taacgcgaat ttt //