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&nbsp;(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
//
