LOCUS       V042718                 7726 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042718
VERSION     V042718
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             964..3216
                     /label="EZH2(NM_004456)"
                     /note="EZH2(NM_004456)"
                     /gene="EZH2"
     CDS             3223..3288
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3326..3550
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3596..4024
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4038..4367
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4434..5228
                     /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    5402..5523
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5572..5588)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5596..5612
                     /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(5620..5650)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5665..5686
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5974..6559)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6730..7590)
                     /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(7591..7695)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgggcc agactgggaa gaaatctgag aagggaccag tttgttggcg gaagcgtgta
     1021 aaatcagagt acatgcgact gagacagctc aagaggttca gacgagctga tgaagtaaag
     1081 agtatgttta gttccaatcg tcagaaaatt ttggaaagaa cggaaatctt aaaccaagaa
     1141 tggaaacagc gaaggataca gcctgtgcac atcctgactt ctgtgagctc attgcgcggg
     1201 actagggagt gttcggtgac cagtgacttg gattttccaa cacaagtcat cccattaaag
     1261 actctgaatg cagttgcttc agtacccata atgtattctt ggtctcccct acagcagaat
     1321 tttatggtgg aagatgaaac tgttttacat aacattcctt atatgggaga tgaagtttta
     1381 gatcaggatg gtactttcat tgaagaacta ataaaaaatt atgatgggaa agtacacggg
     1441 gatagagaat gtgggtttat aaatgatgaa atttttgtgg agttggtgaa tgcccttggt
     1501 caatataatg atgatgacga tgatgatgat ggagacgatc ctgaagaaag agaagaaaag
     1561 cagaaagatc tggaggatca ccgagatgat aaagaaagcc gcccacctcg gaaatttcct
     1621 tctgataaaa tttttgaagc catttcctca atgtttccag ataagggcac agcagaagaa
     1681 ctaaaggaaa aatataaaga actcaccgaa cagcagctcc caggcgcact tcctcctgaa
     1741 tgtaccccca acatagatgg accaaatgct aaatctgttc agagagagca aagcttacac
     1801 tcctttcata cgcttttctg taggcgatgt tttaaatatg actgcttcct acatcgtaag
     1861 tgcaattatt cttttcatgc aacacccaac acttataagc ggaagaacac agaaacagct
     1921 ctagacaaca aaccttgtgg accacagtgt taccagcatt tggagggagc aaaggagttt
     1981 gctgctgctc tcaccgctga gcggataaag accccaccaa aacgtccagg aggccgcaga
     2041 agaggacggc ttcccaataa cagtagcagg cccagcaccc ccaccattaa tgtgctggaa
     2101 tcaaaggata cagacagtga tagggaagca gggactgaaa cggggggaga gaacaatgat
     2161 aaagaagaag aagagaagaa agatgaaact tcgagctcct ctgaagcaaa ttctcggtgt
     2221 caaacaccaa taaagatgaa gccaaatatt gaacctcctg agaatgtgga gtggagtggt
     2281 gctgaagcct caatgtttag agtcctcatt ggcacttact atgacaattt ctgtgccatt
     2341 gctaggttaa ttgggaccaa aacatgtaga caggtgtatg agtttagagt caaagaatct
     2401 agcatcatag ctccagctcc cgctgaggat gtggatactc ctccaaggaa aaagaagagg
     2461 aaacaccggt tgtgggctgc acactgcaga aagatacagc tgaaaaagga cggctcctct
     2521 aaccatgttt acaactatca accctgtgat catccacggc agccttgtga cagttcgtgc
     2581 ccttgtgtga tagcacaaaa tttttgtgaa aagttttgtc aatgtagttc agagtgtcaa
     2641 aaccgctttc cgggatgccg ctgcaaagca cagtgcaaca ccaagcagtg cccgtgctac
     2701 ctggctgtcc gagagtgtga ccctgacctc tgtcttactt gtggagccgc tgaccattgg
     2761 gacagtaaaa atgtgtcctg caagaactgc agtattcagc ggggctccaa aaagcatcta
     2821 ttgctggcac catctgacgt ggcaggctgg gggattttta tcaaagatcc tgtgcagaaa
     2881 aatgaattca tctcagaata ctgtggagag attatttctc aagatgaagc tgacagaaga
     2941 gggaaagtgt atgataaata catgtgcagc tttctgttca acttgaacaa tgattttgtg
     3001 gtggatgcaa cccgcaaggg taacaaaatt cgttttgcaa atcattcggt aaatccaaac
     3061 tgctatgcaa aagttatgat ggttaacggt gatcacagga taggtatttt tgccaagaga
     3121 gccatccaga ctggcgaaga gctgtttttt gattacagat acagccaggc tgatgccctg
     3181 aagtatgtcg gcatcgaaag agaaatggaa atccctctcg aggactacaa agaccatgac
     3241 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     3301 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3361 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3421 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3481 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3541 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4021 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4201 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4321 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4381 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4441 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4501 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4561 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4621 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4681 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4741 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4801 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4861 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4921 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4981 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5041 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5101 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5161 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5221 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5281 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5341 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5401 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5521 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5581 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5641 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5701 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5761 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6481 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6661 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6841 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7321 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7681 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
