LOCUS       V037290                 8248 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037290
VERSION     V037290
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3738
                     /label="ENPP1(NM_006208)"
                     /note="ENPP1(NM_006208)"
                     /gene="ENPP1"
     CDS             3745..3810
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3848..4072
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4118..4546
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4560..4889
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4956..5750
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5924..6045
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6094..6110)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6118..6134
                     /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(6142..6172)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6187..6208
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6496..7081)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7252..8112)
                     /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(8113..8217)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggagc gcgacggctg cgcggggggc gggagccgcg gcggcgaggg cgggcgcgct
     1021 ccccgggagg gcccggcggg gaacggccgc gatcggggcc gcagccacgc tgccgaggcg
     1081 cccggggacc cgcaggcggc cgcgtccttg ctggccccta tggacgtggg ggaggagccg
     1141 ctggagaagg cggcgcgcgc ccgcactgcc aaggacccca acacctataa agtactctcg
     1201 ctggtattgt cagtatgtgt gttaacaaca atacttggtt gtatatttgg gttgaaacca
     1261 agctgtgcca aagaagttaa aagttgcaaa ggtcgctgtt tcgagagaac atttgggaac
     1321 tgtcgctgtg atgctgcctg tgttgagctt ggaaactgct gtttagatta ccaggagacg
     1381 tgcatagaac cagaacatat atggacttgc aacaaattca ggtgtggtga gaaaaggttg
     1441 accagaagcc tctgtgcctg ttcagatgac tgcaaggaca agggcgactg ctgcatcaac
     1501 tacagttctg tgtgtcaagg tgagaaaagt tgggtagaag aaccatgtga gagcattaat
     1561 gagccacagt gcccagcagg gtttgaaacg cctcctaccc tcttattttc tttggatgga
     1621 ttcagggcag aatatttaca cacttggggt ggacttcttc ctgttattag caaactaaaa
     1681 aaatgtggaa catatactaa aaacatgaga ccggtatatc caacaaaaac tttccccaat
     1741 cactacagca ttgtcaccgg attgtatcca gaatctcatg gcataatcga caataaaatg
     1801 tatgatccca aaatgaatgc ttccttttca cttaaaagta aagagaaatt taatcctgag
     1861 tggtacaaag gagaaccaat ttgggtcaca gctaagtatc aaggcctcaa gtctggcaca
     1921 tttttctggc caggatcaga tgtggaaatt aacggaattt tcccagacat ctataaaatg
     1981 tataatggtt cagtaccatt tgaagaaagg attttagctg ttcttcagtg gctacagctt
     2041 cctaaagatg aaagaccaca cttttacact ctgtatttag aagaaccaga ttcttcaggt
     2101 cattcatatg gaccagtcag cagtgaagtc atcaaagcct tgcagagggt tgatggtatg
     2161 gttggtatgc tgatggatgg tctgaaagag ctgaacttgc acagatgcct gaacctcatc
     2221 cttatttcag atcatggcat ggaacaaggc agttgtaaga aatacatata tctgaataaa
     2281 tatttggggg atgttaaaaa tattaaagtt atctatggac ctgcagctcg attgagaccc
     2341 tctgatgtcc cagataaata ctattcattt aactatgaag gcattgcccg aaatctttct
     2401 tgccgggaac caaaccagca cttcaaacct tacctgaaac atttcttacc taagcgtttg
     2461 cactttgcta agagtgatag aattgagccc ttgacattct atttggaccc tcagtggcaa
     2521 cttgcattga atccctcaga aaggaaatat tgtggaagtg gatttcatgg ctctgacaat
     2581 gtattttcaa atatgcaagc cctctttgtt ggctatggac ctggattcaa gcatggcatt
     2641 gaggctgaca cctttgaaaa cattgaagtc tataacttaa tgtgtgattt actgaatttg
     2701 acaccggctc ctaataacgg aactcatgga agtcttaacc accttctaaa gaatcctgtt
     2761 tatacgccaa agcatcccaa agaagtgcac cccctggtac agtgcccctt cacaagaaac
     2821 cccagagata accttggctg ctcatgtaac ccttcgattt tgccgattga ggattttcaa
     2881 acacagttca atctgactgt ggcagaagag aagattatta agcatgaaac tttaccctat
     2941 ggaagaccta gagttctcca gaaggaaaac accatctgtc ttctttccca gcaccagttt
     3001 atgagtggat acagccaaga catcttaatg cccctttgga catcctatac cgtggacaga
     3061 aatgacagtt tctctacgga agacttctcc aactgtctgt accaggactt tagaattcct
     3121 cttagtcctg tccataaatg ttcattttat aaaaataaca ccaaagtgag ttacgggttc
     3181 ctctccccac cacaactaaa taaaaattca agtggaatat attctgaagc tttgcttact
     3241 acaaatatag tgccaatgta ccagagtttt caagttatat ggcgctactt tcatgacacc
     3301 ctactgcgaa agtatgctga agaaagaaat ggtgtcaatg tcgtcagtgg tcctgtgttt
     3361 gactttgatt atgatggacg ttgtgattcc ttagagaatc tgaggcaaaa aagaagagtc
     3421 atccgtaacc aagaaatttt gattccaact cacttcttta ttgtgctaac aagctgtaaa
     3481 gatacatctc agacgccttt gcactgtgaa aacctagaca ccttagcttt cattttgcct
     3541 cacaggactg ataacagcga gagctgtgtg catgggaagc atgactcctc atgggttgaa
     3601 gaattgttaa tgttacacag agcacggatc acagatgttg agcacatcac tggactcagc
     3661 ttctatcaac aaagaaaaga gccagtttca gacattttaa agttgaaaac acatttgcca
     3721 acctttagcc aagaagacct cgaggactac aaagaccatg acggtgatta taaagatcat
     3781 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     3841 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3901 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3961 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4021 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4081 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4141 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4201 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4261 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4321 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4381 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4441 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4501 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4561 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4621 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4681 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4801 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4861 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4921 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4981 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5041 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5101 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5161 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5221 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5281 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5341 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5401 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5461 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5521 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5581 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5641 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5701 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5761 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5821 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5881 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5941 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6001 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6061 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6121 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6181 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6241 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7021 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7081 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7141 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7201 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7261 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7321 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7381 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7441 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7501 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7561 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7621 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7681 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7741 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7801 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7861 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7921 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7981 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8041 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8101 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8161 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8221 atttccccga aaagtgccac ctgacgtc
//
