LOCUS       V021259                 7286 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021259
VERSION     V021259
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             1004..3448
                     /label="Dact1(NM_001190466)"
                     /note="Dact1(NM_001190466)"
                     /gene="Dact1"
     CDS             3452..3481
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3500..3523
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3543..3559)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3570..4192
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4341..4929)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5258..5305
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5537..6331)
                     /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(6366..6723)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6725..6829)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6856..7284
                     /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 tggatccgcc accatgaagc cggacgcagc
     1021 gcgcgagccg gagccgctga gccccggccg gggcgcggag gccgaggggc gctggcgcga
     1081 gaggggcgag gcggacacgg agcggcagcg tacccgcgag cgccaggagg ccacgctggc
     1141 ggggctggcg gagctggggt acctgcggca acgccaagag ctgctggtgc gcggtgcgct
     1201 gcgctgctcc gggaccgtgg ggaccgtcgc gccgcgctcc ggggagctgc ggggagacgc
     1261 ggcgcagcgc agccgcctgg aggagaagtt cctggaagag aacatcttgc tgctgcgaag
     1321 gcagttgaat tgtttgagga gaagagatgc cggtttgttg aatcagttgc aagaacttga
     1381 caagcagata agtgacctga gactggatgt ggagaagaca tctgaagagc acctggagac
     1441 agacagccgg cctagctcag ggttttatga gctgagtgat ggagcttcgg gctccctctc
     1501 taactcctcc aactccgtgt tcagtgagtg tttgtccagt tgccattcca gcacctgctt
     1561 ctgcagcccc ttggaggcgg ccttgaccat ctcagacggt tgccccaaat ctgcagatct
     1621 cataggatgg ttggaatgta aaggcggccc ctgtgaagac caggcctcag ggacagtgtg
     1681 cagttccccc tccacaccac aatttaattc ccttgatgtc attgcagatg tgaatcctaa
     1741 ataccagtgt gatcttgtgt ctaaaaacgg gaatgacgta tatcgctacc ccagtccact
     1801 tcatgctgtg gctgtgcaga gcccaatgtt tctcctttgt ctgacgggca acactctgag
     1861 ggaagaggag gggcttggga gccatgccag cgacatctgc attggatctg aactgaacgc
     1921 caccaaaaca gacaattccc tgccatctcc aagcagtttg tggtccgctt cccatcctgc
     1981 atccagtaag aaaatggatg ggtatatttt gagcctcgtg cagaagaaaa cacaccctgt
     2041 aaggaccaat aaacctagaa ccagtgtgaa cgctgaccct accaagggcc ttctgaggaa
     2101 tggaagtgtg tgtgtcaggg cccctagtgg cgtcccaccg ggcagtagtg tgaactttaa
     2161 gaatacaaaa cagatgtgtt tgcccgctgg gggaataacc tctttggaaa acgggccatt
     2221 ctcccctcct aagcagaggt ccaaagactc aaagacagac cagttagaaa gcaagaggtt
     2281 ggctctgccg gagagctgct cggcaggcgc cgccatggaa ccccaaagca agcatgtgcc
     2341 caaagccgcc aaggcagcct ctcaagagct cacaaggtgt caagccgggc tgggggaatc
     2401 catgaaggaa agcaatcagg cctccgctgt ttctcctaaa acaagtcctg gcagaggccc
     2461 tgtcgccccc gcagagagca aagccctgca gctcccgaaa aagatgtcgc agaagaacag
     2521 cctccaggct gtgcccgcgc tggacaggcc ggccttggac ttcaaaagcg agggctcatc
     2581 tcaaagcctc gaggaagggc atctggtgaa agctcagttc attccggggc agcaggcggc
     2641 cgccaggcct caccgtgcac acaggaaccc gggtgtcgca aggagcgcca ccttgaaggc
     2701 ccgcggccag gcagccatgg aacacggcct gcccaccgtc agggagaaac cgcgggcagc
     2761 aggcaagaag tgccgtttcc cagacgactc ggatacaaat aagaaattca ggaagacctc
     2821 cgccaagggc cggcgcagtg gcgggctgca ggacgctggc cttcccggta gggccctggg
     2881 caccggcggc catcgggcgg gtagcagggc gcacgcgcat ggccgggagc ccgtggtggc
     2941 caaaccgaag cacaagcgaa ccgactaccg gcggtggaaa tcgtcagccg aggtctccta
     3001 cgaagaagcc ctgcggaggg cccggagggc tcgcagggag cacggggctg cctaccgggt
     3061 ggctgtcgcc ctgccttacg ccagccccta tgcctacgtg cccagcgact ccgagtactc
     3121 ggcggagtgc gagtcgctct tccactccac ggtggtggac accagcgagg acgagcagag
     3181 caactacacc accaactgct tcggcgacag cgagtccagc gtgagcgaag gcgacttcgt
     3241 gggcgagagc accaccacca gcgactcaga ggagagcggg ggtttaatct ggtcccagtt
     3301 tgtccagact ctcccgattc aaacggtcac ggccccagac ctccacaccc gtcccacaaa
     3361 aacctttgtc aaaatcaagg cttcgcacaa cctcaagaag aagatcctcc gtttccgctc
     3421 tggctctttg aaactgatga ctaccgttct cgagcagaaa ctcatctcag aagaggatct
     3481 ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa cggccggccg
     3541 cggtcatagc tgtttcctga acagatcccg ggtggcatcc ctgtgacccc tccccagtgc
     3601 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3661 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3721 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3781 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3841 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3901 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3961 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     4021 gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag caggaggacg
     4081 tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc
     4141 ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc
     4201 ggccagcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4261 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4321 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4381 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4441 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4501 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4561 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4621 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4681 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4741 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4801 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4861 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4921 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4981 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5041 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca
     5101 gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg
     5161 ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg
     5221 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc
     5281 gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc
     5341 cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc
     5401 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     5461 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     5521 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     5581 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     5641 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcgatccgcc acacccagcc
     5701 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     5761 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     5821 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     5881 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     5941 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     6001 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     6061 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     6121 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     6181 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     6241 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     6301 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     6361 cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     6421 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     6481 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     6541 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     6601 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6661 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
     6721 caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6841 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6901 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6961 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7021 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7081 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7141 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7201 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7261 tttaacaaaa atttaacgcg aatttt
//
