LOCUS       V034665                 7910 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034665
VERSION     V034665
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3625
                     /label="Enpp1(NM_001308327)"
                     /note="Enpp1(NM_001308327)"
                     /gene="Enpp1"
     polyA_signal    3672..3896
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3942..4370
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4384..4713
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4780..5379
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5553..5674
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5723..5739)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5747..5763
                     /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(5771..5801)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5816..5837
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6125..6710)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6881..7741)
                     /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(7742..7846)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagcgcgacg gcgaccaggc agggcacggg ccccggcacg ggtcagcggg
      961 aaacggtcgc gagctcgagt caccagccgc ggcttcgctg ctcgcgccca tggacctagg
     1021 agaggagccg ctggagaagg cggagcgtgc ccgccctgcc aaagacccca acacctacaa
     1081 agtgctgtcg ctggttttgt cagtatgtgt gctaacaaca attcttggtt gtatatttgg
     1141 gttgaaacca agctgcgcca aagaagtaaa aagttgcaaa ggccgctgct ttgaaaggac
     1201 gttcagcaac tgtcgctgtg atgctgcctg tgtcagcctt gggaactgct gtctggattt
     1261 ccaggagacc tgtgtggaac caacacatat atggacttgc aacaaattca ggtgcggtga
     1321 gaagaggctg tccaggttcg tgtgctcctg tgcagacgac tgcaaaaccc acaatgactg
     1381 ttgcatcaac tacagttctg tgtgccaaga taagaagagt tgggtagaag aaacatgtga
     1441 aagcatcgat accccggagt gtccagcaga gtttgaatca ccccctactc tcttgttttc
     1501 tttggatgga ttcagagctg agtatttgca cacctggggt ggacttcttc ctgtcattag
     1561 caagctgaaa aactgtggaa catacactaa aaacatgagg cctatgtacc ctaccaagac
     1621 gtttcccaat cattacagca tcgtcacagg actttatccg gaatcccatg gcataattga
     1681 taacaagatg tatgatccca aaatgaatgc ttctttctcg cttaaaagta aagagaaatt
     1741 caaccctttg tggtacaaag gacagccgat ttgggtgacc gctaatcatc aggaggtcaa
     1801 gtccggcaca tacttttggc caggatcaga tgtggagatt gacgggattc taccagatat
     1861 ctacaaagtg tataatgggt cagtaccatt tgaagaaagg attttagctg ttcttgagtg
     1921 gctgcagctt cctagccatg aaagaccaca cttttacact ctgtatttag aagaaccaga
     1981 ttcttcaggg cattcacatg gaccagtcag cagcgaggtc atcaaggcct tgcagaaggt
     2041 tgaccgcctg gtcggcatgc tgatggacgg cctgaaggac ctgggcttgg ataaatgcct
     2101 gaacctcatc ctcatttcag atcacggcat ggaacaaggc agctgtaaga agtatgtgta
     2161 cctgaataag tacttggggg atgtgaacaa tgtgaaagtt gtgtatggac ctgctgctcg
     2221 gttgagaccc actgatgttc cagagacata ctattcattt aactatgaag cccttgcaaa
     2281 aaatctttct tgccgggagc caaaccagca tttccggcct tacctgaaac ccttcttacc
     2341 caagcgctta cacttcgcta aaagtgaccg gattgagccc ctgaccttct acctggaccc
     2401 tcagtggcaa cttgcgttga atccatcaga gaggaaatat tgtggaagtg gatttcatgg
     2461 ctctgacaac ttgttttcaa acatgcaagc tctttttatt ggctatggac ctgccttcaa
     2521 gcatggtgct gaagttgact cctttgaaaa tattgaagtc tataacttaa tgtgtgactt
     2581 actgggtttg atcccagctc ccaataatgg aagtcatggc agccttaacc accttctaaa
     2641 gaaacccatt tacaacccaa gtcatcccaa agaagagggc ttcctgtctc agtgtccaat
     2701 caaatcaaca tccaatgacc tcggctgcac atgtgaccct tggattgtgc caatcaagga
     2761 ctttgagaaa cagcttaatc tgaccacaga agatgtagat gacatttacc atatgactgt
     2821 accctacggg aggccccgga ttctactgaa gcagcaccgc gtctgtctac tccagcagca
     2881 acagttcttg accggataca gcctggacct cctcatgccc ctctgggcgt cttacacctt
     2941 cctcagtaat gaccaattct ccagagatga cttttccaac tgtctgtacc aggatcttcg
     3001 gattcccctt agtcctgtgc ataaatgttc ctattacaaa agcaattcta agctaagtta
     3061 tgggttcctc acgccaccga gactaaatag agtttcaaat cacatatact ctgaagcgct
     3121 gcttacatct aatatagtgc caatgtatca gagttttcaa gttatatggc actaccttca
     3181 tgacacccta ctgcaaaggt atgcccatga aaggaacggc atcaatgttg tcagcggtcc
     3241 cgtgtttgac tttgattatg acggacgcta tgattcctta gagatcctga aacaaaacag
     3301 cagagtcatc cgaagccagg aaattctgat tccgactcac ttcttcattg tgctcaccag
     3361 ctgcaagcag ctgtctgaga ctcccttgga gtgctccgcc ttagagtcct cagcttacat
     3421 actgcctcat aggcctgata acattgagag ctgtacgcat gggaagcggg agtcttcatg
     3481 ggtcgaggag ttgctgacct tgcacagagc tcgggtcaca gacgtggaac tcatcactgg
     3541 cctcagcttc taccaggacc gacaagagtc agtttcagaa ctgctgaggt tgaaaacaca
     3601 tttgccaatc ttcagccaag aagactgact cgagtctaga gggcccgttt aaacccgctg
     3661 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3721 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3781 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3841 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3901 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3961 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4021 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4081 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4141 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4201 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4261 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4321 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4381 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4441 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4501 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4561 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4621 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4681 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4741 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgaccgagta caagcccacg
     4801 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
     4861 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     4921 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4981 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5041 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5101 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5161 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5221 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5281 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5341 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag cgggactctg gggttcgaaa
     5401 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5461 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5521 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5641 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5701 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5761 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5821 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6781 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6841 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6901 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6961 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7021 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7081 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7141 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7201 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7261 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7321 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7381 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7441 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7501 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7561 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7621 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7681 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7741 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7801 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7861 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
