LOCUS       V018934                 7694 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018934
VERSION     V018934
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2374
                     /label="NOX1(NM_007052)"
                     /note="NOX1(NM_007052)"
                     /gene="NOX1"
     CDS             2381..2446
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2470..3058
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3124..3172
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3374..3802
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3816..4145
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4212..5006
                     /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    5182..5303
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5352..5368)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5376..5392
                     /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(5400..5430)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5445..5466
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5754..6342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6513..7373)
                     /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(7374..7478)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgggaaa ctgggtggtt aaccactggt tttcagtttt
      721 gtttctggtt gtttggttag ggctgaatgt tttcctgttt gtggatgcct tcctgaaata
      781 tgagaaggcc gacaaatact actacacaag aaaaatcctt gggtcaacat tggcctgtgc
      841 ccgagcgtct gctctctgct tgaattttaa cagcacgctg atcctgcttc ctgtgtgtcg
      901 caatctgctg tccttcctga ggggcacctg ctcattttgc agccgcacac tgagaaagca
      961 attggatcac aacctcacct tccacaagct ggtggcctat atgatctgcc tacatacagc
     1021 tattcacatc attgcacacc tgtttaactt tgactgctat agcagaagcc gacaggccac
     1081 agatggctcc cttgcctcca ttctctccag cctatctcat gatgagaaaa aggggggttc
     1141 ttggctaaat cccatccagt cccgaaacac gacagtggag tatgtgacat tcaccagcat
     1201 tgctggtctc actggagtga tcatgacaat agccttgatt ctcatggtaa cttcagctac
     1261 tgagttcatc cggaggagtt attttgaagt cttctggtat actcaccacc tttttatctt
     1321 ctatatcctt ggcttaggga ttcacggcat tggtggaatt gtccggggtc aaacagagga
     1381 gagcatgaat gagagtcatc ctcgcaagtg tgcagagtct tttgagatgt gggatgatcg
     1441 tgactcccac tgtaggcgcc ctaagtttga agggcatccc cctgagtctt ggaagtggat
     1501 ccttgcaccg gtcattcttt atatctgtga aaggatcctc cggttttacc gctcccagca
     1561 gaaggttgtg attaccaagg ttgttatgca cccatccaaa gttttggaat tgcagatgaa
     1621 caagcgtggc ttcagcatgg aagtggggca gtatatcttt gttaattgcc cctcaatctc
     1681 tctcctggaa tggcatcctt ttactttgac ctctgctcca gaggaagatt tcttctccat
     1741 tcatatccga gcagcagggg actggacaga aaatctcata agggctttcg aacaacaata
     1801 ttcaccaatt cccaggattg aagtggatgg tccctttggc acagccagtg aggatgtttt
     1861 ccagtatgaa gtggctgtgc tggttggagc aggaattggg gtcaccccct ttgcttctat
     1921 cttgaaatcc atctggtaca aattccagtg tgcagaccac aacctcaaaa caaaaaagat
     1981 ctatttctac tggatctgca gggagacagg tgccttttcc tggttcaaca acctgttgac
     2041 ttccctggaa caggagatgg aggaattagg caaagtgggt tttctaaact accgtctctt
     2101 cctcaccgga tgggacagca atattgttgg tcatgcagca ttaaactttg acaaggccac
     2161 tgacatcgtg acaggtctga aacagaaaac ctcctttggg agaccaatgt gggacaatga
     2221 gttttctaca atagctacct cccaccccaa gtctgtagtg ggagttttct tatgtggccc
     2281 tcggactttg gcaaagagcc tgcgcaaatg ctgtcaccga tattccagtc tggatcctag
     2341 aaaggttcaa ttctacttca acaaagaaaa ttttctcgag gactacaaag accatgacgg
     2401 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2461 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2881 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3061 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3121 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3181 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3241 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3301 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3361 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3421 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3481 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3541 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3601 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3661 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3721 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3781 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3961 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4021 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4081 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4141 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4201 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4261 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4321 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4381 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4441 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4501 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4561 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4621 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4681 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4741 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4801 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4861 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4921 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4981 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     5041 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5101 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5161 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5281 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5341 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5401 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5461 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5941 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6181 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7501 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7561 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7621 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7681 agaatctgct tagg
//
