LOCUS       V023269                 8805 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023269
VERSION     V023269
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..3479
                     /label="Dnmt3a(NM_007872)"
                     /note="Dnmt3a(NM_007872)"
                     /gene="Dnmt3a"
     CDS             3492..3557
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3581..4169
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4235..4283
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4485..4913
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4927..5256
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5323..6117
                     /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    6293..6414
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6463..6479)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6487..6503
                     /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(6511..6541)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6556..6577
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6865..7453)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7624..8484)
                     /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(8485..8589)
                     /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 ctcggatccg ccaccatgcc ctccagcggc cccggggaca
      781 ccagcagctc ctctctggag cgggaggatg atcgaaagga aggagaggaa caggaggaga
      841 accgtggcaa ggaagagcgc caggagccca gcgccacggc ccggaaggtg gggaggcctg
      901 gccggaagcg caagcaccca ccggtggaaa gcagtgacac ccccaaggac ccagcagtga
      961 ccaccaagtc tcagcccatg gcccaggact ctggcccctc agatctgcta cccaatggag
     1021 acttggagaa gcggagtgaa ccccaacctg aggaggggag cccagctgca gggcagaagg
     1081 gtggggcccc agctgaagga gagggaactg agaccccacc agaagcctcc agagctgtgg
     1141 agaatggctg ctgtgtgacc aaggaaggcc gtggagcctc tgcaggagag ggcaaagaac
     1201 agaagcagac caacatcgaa tccatgaaaa tggagggctc ccggggccga ctgcgaggtg
     1261 gcttgggctg ggagtccagc ctccgtcagc gacccatgcc aagactcacc ttccaggcag
     1321 gggaccccta ctacatcagc aaacggaaac gggatgagtg gctggcacgt tggaaaaggg
     1381 aggctgagaa gaaagccaag gtaattgcag taatgaatgc tgtggaagag aaccaggcct
     1441 ctggagagtc tcagaaggtg gaggaggcca gccctcctgc tgtgcagcag cccacggacc
     1501 ctgcttctcc gactgtggcc accacccctg agccagtagg aggggatgct ggggacaaga
     1561 atgctaccaa agcagccgac gatgagcctg agtatgagga tggccggggc tttggcattg
     1621 gagagctggt gtgggggaaa cttcggggct tctcctggtg gccaggccga attgtgtctt
     1681 ggtggatgac aggccggagc cgagcagctg aaggcactcg ctgggtcatg tggttcggag
     1741 atggcaagtt ctcagtggtg tgtgtggaga agctcatgcc gctgagctcc ttctgcagtg
     1801 cattccacca ggccacctac aacaagcagc ccatgtaccg caaagccatc tacgaagtcc
     1861 tccaggtggc cagcagccgt gccgggaagc tgtttccagc ttgccatgac agtgatgaaa
     1921 gtgacagtgg caaggctgtg gaagtgcaga acaagcagat gattgaatgg gccctcggtg
     1981 gcttccagcc ctcgggtcct aagggcctgg agccaccaga agaagagaag aatccttaca
     2041 aggaagttta caccgacatg tgggtggagc ctgaagcagc tgcttacgcc ccacccccac
     2101 cagccaagaa acccagaaag agcacaacag agaaacctaa ggtcaaggag atcattgatg
     2161 agcgcacaag ggagcggctg gtgtatgagg tgcgccagaa gtgcagaaac atcgaggaca
     2221 tttgtatctc atgtgggagc ctcaatgtca ccctggagca cccactcttc attggaggca
     2281 tgtgccagaa ctgtaagaac tgcttcttgg agtgtgctta ccagtatgac gacgatgggt
     2341 accagtccta ttgcaccatc tgctgtgggg ggcgtgaagt gctcatgtgt gggaacaaca
     2401 actgctgcag gtgcttttgt gtcgagtgtg tggatctctt ggtggggcca ggagctgctc
     2461 aggcagccat taaggaagac ccctggaact gctacatgtg cgggcataag ggcacctatg
     2521 ggctgctgcg aagacgggaa gactggcctt ctcgactcca gatgttcttt gccaataacc
     2581 atgaccagga atttgacccc ccaaaggttt acccacctgt gccagctgag aagaggaagc
     2641 ccatccgcgt gctgtctctc tttgatggga ttgctacagg gctcctggtg ctgaaggacc
     2701 tgggcatcca agtggaccgc tacattgcct ccgaggtgtg tgaggactcc atcacggtgg
     2761 gcatggtgcg gcaccaggga aagatcatgt acgtcgggga cgtccgcagc gtcacacaga
     2821 agcatatcca ggagtggggc ccattcgacc tggtgattgg aggcagtccc tgcaatgacc
     2881 tctccattgt caaccctgcc cgcaagggac tttatgaggg tactggccgc ctcttctttg
     2941 agttctaccg cctcctgcat gatgcgcggc ccaaggaggg agatgatcgc cccttcttct
     3001 ggctctttga gaatgtggtg gccatgggcg ttagtgacaa gagggacatc tcgcgatttc
     3061 ttgagtctaa ccccgtgatg attgacgcca aagaagtgtc tgctgcacac agggcccgtt
     3121 acttctgggg taaccttcct ggcatgaaca ggcctttggc atccactgtg aatgataagc
     3181 tggagctgca agagtgtctg gagcacggca gaatagccaa gttcagcaaa gtgaggacca
     3241 ttaccaccag gtcaaactct ataaagcagg gcaaagacca gcatttcccc gtcttcatga
     3301 acgagaagga ggacatcctg tggtgcactg aaatggaaag ggtgtttggc ttccccgtcc
     3361 actacacaga cgtctccaac atgagccgct tggcgaggca gagactgctg ggccgatcgt
     3421 ggagcgtgcc ggtcatccgc cacctcttcg ctccgctgaa ggaatatttt gcttgtgtgc
     3481 tcgagtctag agactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3541 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3601 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3661 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3721 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3781 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3841 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3901 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3961 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     4021 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4081 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4141 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4201 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4261 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4321 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4381 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4441 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4621 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4801 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4921 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4981 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5161 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5221 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5281 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5821 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6121 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6181 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6241 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6421 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6481 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6541 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6601 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6661 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     7081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8581 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8641 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8701 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8761 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
