LOCUS V030581 5388 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030581 VERSION V030581 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 1029..1535 /label="Nts(NM_024435)" /note="Nts(NM_024435)" /gene="Nts" CDS 1554..1583 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 1602..1625 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(1645..1661) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 1672..2294 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(2443..3031) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 3360..3407 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(3639..4433) /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(4468..4825) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(4827..4931) /gene="bla" /label="AmpR promoter" rep_origin 4958..5386 /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 gagaggaatg aatctccagc tggtgtgcct gactctcctg gctttcagct 1081 cctggagtct gtgctcagat tcagaagaag atgtgagagc cctggaggca gatctattga 1141 caaacatgca tacatccaag atcagcaaag caagtcctcc gtcttggaaa atgaccttgc 1201 taaatgtttg cagcctcata aataacgtga acagcccggc cgaggaagca ggagacatgc 1261 atgatgacga ccttgttggc aaaaggaaac ttccccttgt tctggatggt tttagcttgg 1321 aagcaatgct gaccatcttc cagctccaga aaatctgccg cagcagggcc tttcaacact 1381 gggagataat ccaggaagat atccttgata acgtcaatga taaaaacgag aaggaagaag 1441 tgataaagag aaaaatccct tatattctga aaaggcagct gtatgaaaat aaacccagaa 1501 ggccctacat tctcaagagg ggttcctact actacacgcg tacgcggccg ctcgagcaga 1561 aactcatctc agaagaggat ctggcagcaa atgatatcct ggattacaag gatgacgacg 1621 ataaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat 1681 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 1741 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 1801 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg 1861 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca 1921 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc 1981 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc 2041 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc 2101 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa 2161 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg 2221 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag 2281 atgcctgttg aattgggtac gcggccagcg gcgagcggta tcagctcact caaaggcggt 2341 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 2401 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 2461 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 2521 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 2581 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 2641 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2701 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2761 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2821 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2881 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2941 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 3001 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 3061 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 3121 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 3181 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc 3241 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca 3301 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg 3361 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt 3421 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa 3481 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc 3541 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg 3601 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg 3661 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg 3721 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga 3781 tagcgatccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc 3841 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc 3901 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc 3961 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt 4021 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca 4081 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc 4141 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct 4201 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca 4261 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc 4321 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc 4381 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac 4441 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc 4501 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa 4561 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg 4621 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct 4681 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct 4741 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt 4801 ccacagctgg ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat 4861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 4921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta 4981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 5041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 5101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 5161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 5221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 5281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 5341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaatttt //