LOCUS       V034755                 5668 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034755
VERSION     V034755
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        27..330
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        331..534
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     intron          672..768
                     /note="modified SV40 intron with splice donor and acceptor
                     sites"
                     /label="SV40 intron"
     CDS             832..855
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             859..2760
                     /label="Trim25(NM_009546)"
                     /note="Trim25(NM_009546)"
                     /gene="Trim25"
     polyA_signal    2807..2941
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3052..3068)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3076..3092
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(3100..3130)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3145..3166
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3454..4042)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4213..5073)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(5074..5178)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     5652..5668
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggactacaag
      841 gacgacgatg acaagcttat ggcggagctg aatcctctgg ccgaggagct gtcctgctcc
      901 gtctgcctgg agctcttcaa ggaacctgtc accactccgt gcggccacaa tttctgcatg
      961 tcatgcctgg atgagacgtg ggtcgtccag ggcccgccgt accgatgccc gcaatgtcgc
     1021 aaagtgtacc aggtgcgccc gcagctgcaa aagaacacgg taatgtgcgc ggtggtggag
     1081 cagttcctgc aggccgagca ggcgcggacc ccggtggacg actggacgcc tcctgcccgc
     1141 ttctccgcct ccagcgcagc cacccaggtg gcctgcgacc actgcctgac ggagatcgca
     1201 gtgaagacgt gcctcgtgtg catggcctct ttctgccagg agcacttgag gcctcacttc
     1261 gacagtccgg ccttccagga tcacccccta cagtcgccca ttcgtgacct tctgcgccgc
     1321 aaatgtaccc agcacaaccg ccttcgggaa ttgttttgtc ccgagcatgg cgagtgcatc
     1381 tgccatatct gcttggtaga gcacaagacc tgctccccta cgaccctaag tcaagccagc
     1441 gccgacctgg agtacaaact gaggaataaa ctcactatca tgcacagcca catcaatggg
     1501 gcaaccaaag cattggagga tgtgagatcc aagcaacagt gtgtgcagga ttctatgaag
     1561 aggaagatgg aacagctgag acaggagtat atggaaatga aggctgtcat cgacgcagca
     1621 gagaccagct ccttgcgcaa gttaaaggag gaggagaaga gagtctacgg aaaattcgac
     1681 accatctacc aggttctcgt caagaagaag agtgagatgc agaaactgaa ggcagaggtt
     1741 gagctcatta tggacaaagg ggatgagttt gagtttctgg agaaagctgc aaaattgcaa
     1801 ggagagtcaa caaaaccagt ctacatccct aagatagatc tggatcatga tttgataatg
     1861 ggaatctacc aaggggcggc ggacctcaag agtgaactga agcactccat taagaagcta
     1921 cagaagaagt ctgaggagca caatggctca ggtaacaagg gagaccagac acagtccact
     1981 tttaaacccg tacaaccttc gaagaagacc attcaagaaa agaagaccaa gaaaacccct
     2041 gttgctcccg gccctccctc ccacttctca cctaacaagt tgcccacctt tggagctcct
     2101 ggccaatcac tggattcaaa agcaacttcc cctgatgccg cacccaaagc cagtgctgca
     2161 cagccagact ctgtaggagt caaggctaag gtgctagaaa acttcttaac caagtccaga
     2221 acggagcttc tggagtattt cgtgaaagtc atcttcgact acaataccgc ccacaacaaa
     2281 gtgtctctgt caaacaagta caccaccgcc tctgtgtccg atgggctcca gcactaccgg
     2341 tcccatcccc agaggttcac ctactgctct caggtcctgg ggctgcactg ctacaagaac
     2401 ggcattcact actgggaggt ggaactacag aagaacaact tctgcggcgt aggcatctgc
     2461 tacggcagca tggaacggca gggccccgag agccggctgg gccgcaaccc caactcatgg
     2521 tgtgtggagt ggttcaataa caagatctct gcctggcaca acaacgtgga gaagactctg
     2581 ccctccacca aggccactcg agtcggcgtg ctgctcaact gtgaccacgg cttcgtcatt
     2641 ttctttgcgg tcaccgaaaa ggtgcacctg atgtacaagt tcaaggtgga cttcaccgaa
     2701 gctctgtacc cagccttctg ggtgttctcg gccggtacca ccctctctat ctgctccaaa
     2761 taggaattcg gtcgaccgag atctctcgag gtaccgcggc cgcggggatc cagacatgat
     2821 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2881 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2941 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3001 ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat ggtcatagct
     3061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5221 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5281 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5341 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5401 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5461 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5521 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5581 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5641 gtcacgacgt tgtaaaacga cggccagt
//
