LOCUS       V022535                 7148 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022535
VERSION     V022535
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..1828
                     /label="GJA1(NM_000165)"
                     /note="GJA1(NM_000165)"
                     /gene="GJA1"
     CDS             1835..1900
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1924..2512
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2578..2626
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2828..3256
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3270..3599
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3666..4460
                     /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    4636..4757
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4806..4822)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4830..4846
                     /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(4854..4884)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4899..4920
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5967..6827)
                     /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(6828..6932)
                     /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 ccatgggtga ctggagcgcc ttaggcaaac tccttgacaa
      721 ggttcaagcc tactcaactg ctggagggaa ggtgtggctg tcagtacttt tcattttccg
      781 aatcctgctg ctggggacag cggttgagtc agcctgggga gatgagcagt ctgcctttcg
      841 ttgtaacact cagcaacctg gttgtgaaaa tgtctgctat gacaagtctt tcccaatctc
      901 tcatgtgcgc ttctgggtcc tgcagatcat atttgtgtct gtacccacac tcttgtacct
      961 ggctcatgtg ttctatgtga tgcgaaagga agagaaactg aacaagaaag aggaagaact
     1021 caaggttgcc caaactgatg gtgtcaatgt ggacatgcac ttgaagcaga ttgagataaa
     1081 gaagttcaag tacggtattg aagagcatgg taaggtgaaa atgcgagggg ggttgctgcg
     1141 aacctacatc atcagtatcc tcttcaagtc tatctttgag gtggccttct tgctgatcca
     1201 gtggtacatc tatggattca gcttgagtgc tgtttacact tgcaaaagag atccctgccc
     1261 acatcaggtg gactgtttcc tctctcgccc cacggagaaa accatcttca tcatcttcat
     1321 gctggtggtg tccttggtgt ccctggcctt gaatatcatt gaactcttct atgttttctt
     1381 caagggcgtt aaggatcggg ttaagggaaa gagcgaccct taccatgcga ccagtggtgc
     1441 gctgagccct gccaaagact gtgggtctca aaaatatgct tatttcaatg gctgctcctc
     1501 accaaccgct cccctctcgc ctatgtctcc tcctgggtac aagctggtta ctggcgacag
     1561 aaacaattct tcttgccgca attacaacaa gcaagcaagt gagcaaaact gggctaatta
     1621 cagtgcagaa caaaatcgaa tggggcaggc gggaagcacc atctctaact cccatgcaca
     1681 gccttttgat ttccccgatg ataaccagaa ttctaaaaaa ctagctgctg gacatgaatt
     1741 acagccacta gccattgtgg accagcgacc ttcaagcaga gccagcagtc gtgccagcag
     1801 cagacctcgg cctgatgacc tggagatcct cgaggactac aaagaccatg acggtgatta
     1861 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     1921 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     1981 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2041 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2101 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2161 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2221 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2281 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     2341 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2401 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2461 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     2521 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     2581 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     2641 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     2701 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     2761 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     2821 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3181 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3421 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3481 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3541 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3601 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3661 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3721 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3781 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3841 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3901 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3961 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4021 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4081 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4141 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4201 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4261 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4321 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4381 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4441 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     4501 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4561 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4621 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4741 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6961 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     7021 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7081 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     7141 tgcttagg
//
