LOCUS       V023472                10819 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023472
VERSION     V023472
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             932..6304
                     /label="SCN11A(NM_014139)"
                     /note="SCN11A(NM_014139)"
                     /gene="SCN11A"
     CDS             6320..6385
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    6414..6638
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6684..7112
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7126..7455
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7522..8316
                     /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    8492..8613
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8662..8678)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8686..8702
                     /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(8710..8740)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8755..8776
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(9064..9652)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9823..10683)
                     /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(10684..10788)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccgccac catggatgac agatgctacc cagtaatctt
      961 tccagatgag cggaatttcc gccccttcac ttccgactct ctggctgcaa ttgagaagcg
     1021 gattgccatc caaaaggaga aaaagaagtc taaagaccag acaggagaag taccccagcc
     1081 tcggcctcag cttgacctaa aggcctccag gaagttgccc aagctctatg gcgacattcc
     1141 tcgtgagctc ataggaaagc ctctggaaga cttggaccca ttctaccgaa atcataagac
     1201 atttatggtg ttaaacagaa agaggacaat ctaccgcttc agtgccaagc atgccttgtt
     1261 catttttggg cctttcaatt caatcagaag tttagccatt agagtctcag tccattcatt
     1321 gttcagcatg ttcattatcg gcaccgttat catcaactgc gtgttcatgg ctacagggcc
     1381 tgctaaaaac agcaacagta acaatactga cattgcagag tgtgtcttca ctgggattta
     1441 tatttttgaa gctttgatta aaatattggc aagaggtttc attctggatg agttttcttt
     1501 ccttcgagat ccatggaact ggctggactc cattgtcatt ggaatagcga ttgtgtcata
     1561 tattccagga atcaccatca aactattgcc cctgcgtacc ttccgtgtgt tcagagcttt
     1621 gaaagcaatt tcagtagttt cacgtctgaa ggtcatcgtg ggggccttgc tacgctctgt
     1681 gaagaagctg gtcaacgtga ttatcctcac cttcttttgc ctcagcatct ttgccctggt
     1741 aggtcagcag ctcttcatgg gaagtctgaa cctgaaatgc atctcgaggg actgtaaaaa
     1801 tatcagtaac ccggaagctt atgaccattg ctttgaaaag aaagaaaatt cacctgaatt
     1861 caaaatgtgt ggcatctgga tgggtaacag tgcctgttcc atacaatatg aatgtaagca
     1921 caccaaaatt aatcctgact ataattatac gaattttgac aactttggct ggtcttttct
     1981 tgccatgttc cggctgatga cccaagattc ctgggagaag ctttatcaac agaccctgcg
     2041 tactactggg ctctactcag tcttcttctt cattgtggtc attttcctgg gctccttcta
     2101 cctgattaac ttaaccctgg ctgttgttac catggcatat gaggagcaga acaagaatgt
     2161 agctgcagag atagaggcca aggaaaagat gtttcaggaa gcccagcagc tgttaaagga
     2221 ggaaaaggag gctctggttg ccatgggaat tgacagaagt tcacttactt cccttgaaac
     2281 atcatatttt accccaaaaa agagaaagct ctttggtaat aagaaaagga agtccttctt
     2341 tttgagagag tctgggaaag accagcctcc tgggtcagat tctgatgaag attgccaaaa
     2401 aaagccacag ctcctagagc aaaccaaacg actgtcccag aatctatcac tggaccactt
     2461 tgatgagcat ggagatcctc tccaaaggca gagagcactg agtgctgtca gcatcctcac
     2521 catcaccatg aaggaacaag aaaaatcaca agagccttgt ctcccttgtg gagaaaacct
     2581 ggcatccaag tacctcgtgt ggaactgttg cccccagtgg ctgtgcgtta agaaggtcct
     2641 gagaactgtg atgactgacc cgtttactga gctggccatc accatctgca tcatcatcaa
     2701 cactgtcttc ttggccatgg agcatcacaa gatggaggcc agttttgaga agatgttgaa
     2761 tatagggaat ttggttttca ctagcatttt tatagcagaa atgtgcctaa aaatcattgc
     2821 gctcgatccc taccactact ttcgccgagg ctggaacatt tttgacagca ttgttgctct
     2881 tctgagtttt gcagatgtaa tgaactgtgt acttcaaaag agaagctggc cattcttgcg
     2941 ttccttcaga gtgctcaggg tcttcaagtt agccaaatcc tggccaactt tgaacacact
     3001 aattaagata atcggcaact ctgtcggagc ccttggaagc ctgactgtgg tcctggtcat
     3061 tgtgatcttt attttctcag tagttggcat gcagcttttt ggccgtagct tcaattccca
     3121 aaagagtcca aaactctgta acccgacagg cccgacagtc tcatgtttac ggcactggca
     3181 catgggggat ttctggcact ccttcctagt ggtattccgc atcctctgcg gggaatggat
     3241 cgaaaatatg tgggaatgta tgcaagaagc gaatgcatca tcatcattgt gtgttattgt
     3301 cttcatattg atcacggtga taggaaaact tgtggtgctc aacctcttca ttgccttact
     3361 gctcaattcc tttagcaatg aggaaagaaa tggaaactta gaaggagagg ccaggaaaac
     3421 taaagtccag ttagcactgg atcgattccg ccgggctttt tgttttgtga gacacactct
     3481 tgagcatttc tgtcacaagt ggtgcaggaa gcaaaactta ccacagcaaa aagaggtggc
     3541 aggaggctgt gctgcacaaa gcaaagacat cattcccctg gtcatggaga tgaaaagggg
     3601 ctcagagacc caggaggagc ttggtatact aacctctgta ccaaagaccc tgggcgtcag
     3661 gcatgattgg acttggttgg caccacttgc ggaggaggaa gatgacgttg aattttctgg
     3721 tgaagataat gcacagcgca tcacacaacc tgagcctgaa caacaggcct atgagctcca
     3781 tcaggagaac aagaagccca cgagccagag agttcaaagt gtggaaattg acatgttctc
     3841 tgaagatgag cctcatctga ccatacagga tccccgaaag aagtctgatg ttaccagtat
     3901 actatcagaa tgtagcacca ttgatcttca ggatggcttt ggatggttac ctgagatggt
     3961 tcccaaaaag caaccagaga gatgtttgcc caaaggcttt ggttgctgct ttccatgctg
     4021 tagcgtggac aagagaaagc ctccctgggt catttggtgg aacctgcgga aaacctgcta
     4081 ccaaatagtg aaacacagct ggtttgagag ctttattatc tttgtgattc tgctgagcag
     4141 tggggcactg atatttgaag atgttcacct tgagaaccaa cccaaaatcc aagaattact
     4201 aaattgtact gacattattt ttacacatat ttttatcctg gagatggtac taaaatgggt
     4261 agccttcgga tttggaaagt atttcaccag tgcctggtgc tgccttgatt tcatcattgt
     4321 gattgtctct gtgaccaccc tcattaactt aatggaattg aagtccttcc ggactctacg
     4381 agcactgagg cctcttcgtg cgctgtccca gtttgaagga atgaaggtgg tggtcaatgc
     4441 tctcataggt gccatacctg ccattctgaa tgttttgctt gtctgcctca ttttctggct
     4501 cgtattttgt attctgggag tatacttctt ttctggaaaa tttgggaaat gcattaatgg
     4561 aacagactca gttataaatt ataccatcat tacaaataaa agtcaatgtg aaagtggcaa
     4621 tttctcttgg atcaaccaga aagtcaactt tgacaatgtg ggaaatgctt acctcgctct
     4681 gctgcaagtg gcaacattta agggctggat ggatattata tatgcagctg ttgattccac
     4741 agagaaagaa caacagccag agtttgagag caattcactc ggttacattt acttcgtagt
     4801 ctttatcatc tttggctcat tcttcactct gaatctcttc attggcgtta tcattgacaa
     4861 cttcaaccaa cagcagaaaa agttaggtgg ccaagacatt tttatgacag aagaacagaa
     4921 gaaatactat aatgcaatga aaaaattagg atccaaaaaa cctcaaaaac ccattccacg
     4981 gcctctgaac aaatgtcaag gtctcgtgtt cgacatagtc acaagccaga tctttgacat
     5041 catcatcata agtctcatta tcctaaacat gattagcatg atggctgaat catacaacca
     5101 acccaaagcc atgaaatcca tccttgacca tctcaactgg gtctttgtgg tcatctttac
     5161 gttagaatgt ctcatcaaaa tctttgcttt gaggcaatac tacttcacca atggctggaa
     5221 tttatttgac tgtgtggtcg tgcttctttc cattgttagt acaatgattt ctaccttgga
     5281 aaatcaggag cacattcctt tccctccgac gctcttcaga attgtccgct tggctcggat
     5341 tggccgaatc ctgaggcttg tccgggctgc acgaggaatc aggactctcc tctttgctct
     5401 gatgatgtcg cttccttctc tgttcaacat tggtcttcta ctctttctga ttatgtttat
     5461 ctatgccatt ctgggtatga actggttttc caaagtgaat ccagagtctg gaatcgatga
     5521 catattcaac ttcaagactt ttgccagcag catgctctgt ctcttccaga taagcacatc
     5581 agcaggttgg gattccctgc tcagccccat gctgcgatca aaagaatcat gtaactcttc
     5641 ctcagaaaac tgccacctcc ctggcatagc cacatcctac tttgtcagtt acattatcat
     5701 ctcctttctc attgttgtca acatgtacat tgctgtgatt ttagagaact tcaatacagc
     5761 cactgaagaa agtgaggacc ctttgggtga agatgacttt gacatatttt atgaagtgtg
     5821 ggaaaagttt gacccagaag caacacaatt tatcaaatat tctgcccttt ctgactttgc
     5881 tgatgccttg cctgagcctt tgcgtgtcgc aaagccaaat aaatatcaat ttctagtaat
     5941 ggacttgccc atggtgagtg aagatcgcct ccactgcatg gatattcttt tcgccttcac
     6001 cgctagggta ctcggtggct ctgatggcct agatagtatg aaagcaatga tggaagagaa
     6061 gttcatggaa gccaatcctc tcaagaagtt gtatgaaccc atagtcacca ccaccaagag
     6121 aaaggaagag gaaagaggtg ctgctattat tcaaaaggcc tttcgaaagt acatgatgaa
     6181 ggtgaccaag ggtgaccaag gtgaccaaaa tgacttggaa aacgggcctc attcaccact
     6241 ccagactctt tgcaatggag acttgtctag ctttggggtg gccaagggca aggtccactg
     6301 tgactctaga gggcccttcg actacaaaga ccatgacggt gattataaag atcatgacat
     6361 cgactacaag gatgacgatg acaagtgagt ttaaacccgc tgatcagcct cgactgtgcc
     6421 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6481 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6541 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6601 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     6661 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     6721 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6781 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6841 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6901 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6961 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7021 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7081 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     7141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7201 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     7261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     7321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7381 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7441 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     7501 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     7561 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     7621 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     7681 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     7741 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     7801 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     7861 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     7921 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     7981 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     8041 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     8101 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     8161 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     8221 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     8281 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     8341 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     8401 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     8461 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     8521 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     8581 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     8641 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8701 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8761 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8821 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8881 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8941 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     9001 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     9061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     9121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     9181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     9241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    10021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    10081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    10141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    10201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10801 aaaagtgcca cctgacgtc
//
