LOCUS       V027309                 5486 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027309
VERSION     V027309
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             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..1797
                     /label="REC114(NM_001042367)"
                     /note="REC114(NM_001042367)"
                     /gene="REC114"
     polyA_signal    1827..2449
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        2478..2807
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(2846..2864)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(2878..2894)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    2902..2918
                     /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(2926..2956)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    2971..2992
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3280..3868)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4039..4899)
                     /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(4900..5004)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5031..5459
                     /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 attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tggcggaggc aggaaaagtg
     1021 cccttgagcc tcgggcttac cggaggagaa gcggcagagt ggcctctgca gcggtacgcc
     1081 cgctgcatac cctcaaacac cagagaccca cctgggccat gcctggaagc tgggacagcc
     1141 ccctgcccca catggaaggt ttttgattcc aatgaagaat ctggatatct tgttctcacc
     1201 atagttatat caggtcattt cttcattttc caaggacaga cactactgga agggttttca
     1261 ctcattggta gcaaggactg gttgaagatt gtaagacgcg tggattgtct gttgtttgga
     1321 acaacgataa aggacaagag tcgcctgttt cgagtacagt tcagtggaga gtcaaaggag
     1381 caggcgctgg aacactgctg cagttgtgtt cagaagctgg cacaatacat aaccgtgcag
     1441 gtgcctgatg gaaacatcca ggagcttcag ctgattcctg gcccacccag ggcaactgaa
     1501 agtcaaggga aggattctgc aaagagtgtc ccacggcagc ctggatccca ccagcactca
     1561 gaacaacagc aagtgtgtgt aacagcgggc acaggcgctc cagacggaag gacctcactg
     1621 acgcagttag ctcagactct tctggcatcg gaggagctgc cccatgtcta tgaacaatct
     1681 gcatggggtg cagaagagtt aggccccttc ctacgtttgt gccttatgga tcagaatttc
     1741 ccagcatttg tggaagaggt agaaaaggaa ctgaaaaagc tggcgggttt gagaaattaa
     1801 ggtaccagtc gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc
     1861 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     1921 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     1981 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     2041 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     2101 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     2161 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     2221 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     2281 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
     2341 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
     2401 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
     2461 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2521 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2641 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2701 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2761 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg
     2821 aactgaaaaa ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc
     2881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2941 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3721 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4981 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     5041 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5101 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5161 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5221 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5281 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5341 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5401 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5461 taacaaaata ttaacgctta caattt
//
