LOCUS       V034146                 8808 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034146
VERSION     V034146
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3488
                     /label="KCNB2(NM_004770)"
                     /note="KCNB2(NM_004770)"
                     /gene="KCNB2"
     CDS             3495..3560
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3584..4172
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4238..4286
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4488..4916
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4930..5259
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5326..6120
                     /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    6296..6417
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6466..6482)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6490..6506
                     /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(6514..6544)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6559..6580
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6868..7456)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7627..8487)
                     /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(8488..8592)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agaaaaggct cccccgggct
      781 taaacaggaa gacttcaagg tcgacacttt cccttcctcc agagcctgtg gacattatcc
      841 ggagcaaaac atgctccagg agagttaaga tcaatgtggg gggcctcaac cacgaagtcc
      901 tgtggagaac gctggacagg ctgcccagga cgcgcctggg gaagcttcga gactgcaaca
      961 cacacgagag cctcctggaa gtgtgcgacg actataatct gaacgagaac gagtatttct
     1021 ttgatcggca tccaggagcc ttcacttcca ttttaaattt ctaccggacc gggaaactcc
     1081 atatgatgga agaaatgtgt gcactttcgt ttggccaaga acttgattac tgggggattg
     1141 atgagatcta cttggagtcc tgctgccagg ccagatatca tcaaaaaaaa gaacaaatga
     1201 acgaagaact gaggcgagag gcagagacta tgcgagagcg agaaggagaa gagtttgata
     1261 atacctgctg ccctgataaa aggaagaaac tgtgggactt gctggagaaa cctaactcat
     1321 cagtggctgc aaagatcctg gccatcgtgt ctatcctgtt cattgtgctt tccaccattg
     1381 ctttgtctct caatacgctg ccggagctgc aggaaacgga cgaatttgga caactcaatg
     1441 acaaccgcca attagcacac gtggaggctg tgtgtattgc atggtttacc atggagtacc
     1501 ttttgcgatt cttatcctca ccaaataaat ggaagttctt caaaggccca ctgaatgtca
     1561 ttgatttgct ggccatcttg ccgtactatg tcaccatttt tctgacggag tccaacaaga
     1621 gcgtgctgca gttccaaaac gtgaggcgcg tggtccagat cttccgaatc atgcgcatcc
     1681 tcaggatcct gaaactcgcc aggcattcga caggcctgca gtctctgggt ttcaccctta
     1741 ggcggagtta caatgaattg ggcttgttga tattgtttct ggccatgggg ataatgatat
     1801 tttccagcct ggtatttttt gctgagaagg atgaagatgc taccaagttc accagtatcc
     1861 ctgcatcatt ttggtgggcc accatcacca tgaccactgt tggctatggt gacatttacc
     1921 ctaaaacatt actagggaaa attgtgggag gtctgtgctg tattgctggg gttctggtta
     1981 ttgcccttcc tatcccaatt attgtgaaca atttttctga gttttacaag gagcagaaac
     2041 gccaagagaa agcaattaaa aggagggagg ctcttgagcg ggccaaaagg aacggaagca
     2101 tcgtttctat gaacttaaaa gatgccttcg ctcgaagtat ggaactgata gatgtggctg
     2161 ttgagaaggc cggagagtcc gccaacacaa aggactccgc cgacgataat cacctgtcgc
     2221 caagccggtg gaagtgggcc aggaaggctc tgtcggaaac aagctccaac aagtctttcg
     2281 agaataagta ccaggaggtt agccaaaaag actcccacga gcagctgaac aacacgtctt
     2341 cctccagccc acagcatctg agtgcccaga aactggagat gctatacaat gaaatcacca
     2401 agacacagcc tcattctcac ccaaacccag actgccaaga aaagcctgag aggccatctg
     2461 catatgaaga agagattgaa atggaagaag tggtgtgtcc acaggagcag ctggccgtgg
     2521 cacagaccga ggtcattgtg gacatgaaga gcacctccag catcgacagc ttcaccagct
     2581 gtgccaccga cttcacagag acagagagat cgccgctgcc gccgccctcc gcctctcact
     2641 tgcagatgaa gttcccaacc gacctcccag ggacagaaga gcaccaaaga gctaggggcc
     2701 ccccgtttct aactctatcc agagagaaag gacctgctgc cagggatggc acgctggagt
     2761 atgccccagt tgacataact gtgaacctcg atgccagtgg ctcccagtgt gggctacata
     2821 gtcctttgca gtctgacaat gccaccgaca gtcctaagag ctctctaaaa ggcagcaacc
     2881 cactaaagtc cagatccctc aaagtgaact ttaaggaaaa tagaggcagt gcaccacaga
     2941 ccccgcccag cacagccagg ccactgccag tcaccacagc tgacttttcg ctcactaccc
     3001 cgcagcacat cagtaccatc ctcttagaag aaaccccctc ccagggagac agacccttgc
     3061 tgggcactga ggtttcagcg ccttgtcagg gaccttccaa agggctgtcc cccaggtttc
     3121 ccaagcagaa actgttccct ttctcttcaa gagagaggag gagcttcact gaaatagata
     3181 ctggtgacga cgaagacttc ttagagctcc caggggcaag ggaggagaag caggtggact
     3241 ccagcccaaa ttgctttgca gataagccta gtgatgggag agacccttta agagaagagg
     3301 gcagtgtggg ctcttcctcc ccgcaggaca caggtcacaa ctgtaggcaa gacatttacc
     3361 atgctgtgag tgaagtcaaa aaggacagta gtcaagaagg gtgcaagatg gaaaatcact
     3421 tgtttgcccc agaaattcat tccaacccag gagacacagg ttattgtccc acacgtgaaa
     3481 ccagcatggg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     3541 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     3601 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3661 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3721 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3781 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3841 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3901 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3961 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4021 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4081 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4141 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     4201 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4261 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4321 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4381 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4441 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     4501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4621 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     4681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5161 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5281 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5341 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5401 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5461 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5521 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5581 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5641 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5701 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5761 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5821 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5881 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5941 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6001 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6061 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6121 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6181 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6241 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6301 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6361 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6421 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6481 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6541 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6601 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6661 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7501 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8521 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8581 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     8641 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     8701 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     8761 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
