LOCUS       V035613                 6416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035613
VERSION     V035613
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             962..1675
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1712..2746
                     /label="OGG1(NM_002542)"
                     /note="OGG1(NM_002542)"
                     /gene="OGG1"
     polyA_signal    2757..3379
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3408..3737
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(3776..3794)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3808..3824)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3832..3848
                     /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(3856..3886)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3901..3922
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4210..4798)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4969..5829)
                     /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(5830..5934)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5961..6389
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1141 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1381 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1441 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1561 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg
     1681 cggagggagt ggaggagggg gaagcgaatt catgcctgcc cgcgcgcttc tgcccaggcg
     1741 catggggcat cgtactctag cctccactcc tgccctgtgg gcctccatcc cgtgccctcg
     1801 ctctgagctg cgcctggacc tggttctgcc ttctggacaa tctttccggt ggagggagca
     1861 aagtcctgca cactggagtg gtgtactagc ggatcaagta tggacactga ctcagactga
     1921 ggagcagctc cactgcactg tgtaccgagg agacaagagc caggctagca ggcccacacc
     1981 agacgagctg gaggccgtgc gcaagtactt ccagctagat gttaccctgg ctcaactgta
     2041 tcaccactgg ggttccgtgg actcccactt ccaagaggtg gctcagaaat tccaaggtgt
     2101 gcgactgctg cgacaagacc ccatcgaatg ccttttctct tttatctgtt cctccaacaa
     2161 caacatcgcc cgcatcactg gcatggtgga gcggctgtgc caggcttttg gacctcggct
     2221 catccagctt gatgatgtca cctaccatgg cttccccagc ctgcaggccc tggctgggcc
     2281 agaggtggag gctcatctca ggaagctggg cctgggctat cgtgcccgtt acgtgagtgc
     2341 cagtgcccga gccatcctgg aagaacaggg cgggctagcc tggctgcagc agctacgaga
     2401 gtcctcatat gaggaggccc acaaggccct ctgcatcctg cctggagtgg gcaccaaggt
     2461 ggctgactgc atctgcctga tggccctaga caagccccag gctgtgcccg tggatgtcca
     2521 tatgtggcac attgcccaac gtgactacag ctggcaccct accacgtccc aggcgaaggg
     2581 accgagcccc cagaccaaca aggaactggg aaactttttc cggagcctgt ggggacctta
     2641 tgctggctgg gcccaagcgg tgctgttcag tgccgacctg cgccaatccc gccatgctca
     2701 ggagccacca gcaaagcgca gaaagggttc caaagggccg gaaggctagg gatcccgggt
     2761 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2821 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2881 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2941 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3001 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3061 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3121 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3181 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3241 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     3301 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     3361 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3601 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3661 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3721 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
     3781 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     3841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5941 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6001 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6061 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6121 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6181 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6241 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6301 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6361 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//
