LOCUS       V029751                 6948 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029751
VERSION     V029751
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..3095
                     /label="Carf(NM_139150)"
                     /note="Carf(NM_139150)"
                     /gene="Carf"
     CDS             3114..3143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3162..3185
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3205..3221)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3232..3854
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4003..4591)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4920..4967
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5199..5993)
                     /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(6028..6385)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6387..6491)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6518..6946
                     /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 ggagcagcgt cattccttag cgggcaactg tgacgatggt gagaagtcag
     1081 aaagagaggc tcagggattc gagcatcgga cctgtatgga ctctggggat ccttcctttg
     1141 ggcaaaatga tccccctacc attttgccca ttactgctcc taaaacatat gattcactca
     1201 catcacagga tatgccagag acactgactg acacacaggc ccttcatgaa gagcaatttc
     1261 atcttttgga ccaaaatggg cagcctattc aatatgatct tcagtctttg ggaaattcta
     1321 ctgcacaaat gacagggatg gagcaagtgg ccatacctca ggagcccttt gcagatgaac
     1381 atagccctca agatatctct gaagagaaac ccactggtgt aagagcagat gttctcgaag
     1441 acagcaccag caattacggc cttcgctcac cagcatccct cgtgctaccc aaaaaggcag
     1501 gagccagact tgttgaagaa cctttgctgg ctcctctcca gccactttct tgtaacacac
     1561 ccatgtgggc ctgccgtctt aggagttgtg agaaaattgg agattcctac cgtggctact
     1621 gtgtaagtga gactgaatta gaaagtgtcc taacttttca caagcagcaa acacagactg
     1681 tctggggtac tcgccagtct ccaagcccag ccaaacctgc cacacgcttg atgtggaagt
     1741 ctcagtatgt cccatatgac ggaatcccat ttgtcaatgc aggaagtcga gctgtggtaa
     1801 tggagtgtca gtatggacca aggagaaaag gttttcagtt aaaaaaaatc agtgagcaag
     1861 aaagtaggtc ttgtcatctc tataaagcta cttgtccagc caggatatac attaaaaagg
     1921 tacagaaatt tcctgaatat agagtaccta cagaccccca aattgacagg aagataatca
     1981 gactggagca ggagaaagcc ttcactatgc tgaagaagaa cctgatggat gctggtggtg
     2041 tacttcgatg gtatgtgcag ttacctacac agcaagctca tcagtatcat gaatcagaga
     2101 ctcctggtgt ccctttgtca ccgtcacctt ttcctatgtc tcctcttgaa gaagaggaag
     2161 ctatagtcag agatgagaac tgtgcgttgc cctctcgtct ccacccgcaa gtagcgcata
     2221 agattcagga actagtgtca cagggagtag gacaagtgta tgcagtaagg aagcagctaa
     2281 gaaaatttgt ggaaagagaa ctgttcaaac ctgatgagat acctgaaaga cataatttat
     2341 cttattttcc aactgtaaat gatataaaaa atcatatcca tgaggtacag aaatccttga
     2401 ggactggaga tgtggtatat aactcagaga ttattccagc aacgcttcag tggacgacag
     2461 atagtgggaa catcctcaga gagactgtaa cagttacatt tgcagaagga aatttactag
     2521 gagaaccgat tcccagcaaa atgggaacaa gtcagacaca gactgctgtg tctccagagc
     2581 cactttcttc atttccaccc aaaatattta cacactttca ggctttgaag ttacagccaa
     2641 gactctcctc tcctgatggc tcacaagctt tggtatcagt agatagccat gcatcctcta
     2701 gtcctcctgg ccttgtggat actgtaggaa atgctgaagt ggataaccac tctgtactgc
     2761 ttggtcagag tcaaaaccct gggacagata catgcctaac ccaagacaac agcacctcct
     2821 cctccacggg ccacctccca gaatcagttc cgaatccagt tgcagaggat cagctgctgg
     2881 aaggtgaaga tgttgaggat gcagggaatc cggaaggaag tgtaaatagg actctgttgg
     2941 gagatgtaca aaccgttcca atacagatca tagacagccg tccagtgctt gttgaagaaa
     3001 gtctgtctaa gaaccaagtt aaacaagaaa ccaatgagcc aacactgtct acagaagcaa
     3061 aaactttcct ggactgtaaa aaaatatctg ctacaacgcg tacgcggccg ctcgagcaga
     3121 aactcatctc agaagaggat ctggcagcaa atgatatcct ggattacaag gatgacgacg
     3181 ataaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc cgggtggcat
     3241 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3301 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3361 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3421 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3481 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3541 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3601 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3661 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     3721 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     3781 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     3841 atgcctgttg aattgggtac gcggccagcg gcgagcggta tcagctcact caaaggcggt
     3901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4741 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc
     4801 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca
     4861 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg
     4921 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     4981 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     5041 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     5101 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     5161 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     5221 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     5281 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     5341 tagcgatccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     5401 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     5461 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     5521 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     5581 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     5641 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     5701 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     5761 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     5821 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     5881 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     5941 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     6001 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     6061 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     6121 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     6181 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6241 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     6301 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     6361 ccacagctgg ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat
     6421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6481 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     6541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaatttt
//
