LOCUS       V043578                 7311 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043578
VERSION     V043578
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..1991
                     /label="Suv39h1(NM_011514)"
                     /note="Suv39h1(NM_011514)"
                     /gene="Suv39h1"
     CDS             1998..2063
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2087..2675
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2741..2789
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2991..3419
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3433..3762
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3829..4623
                     /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    4799..4920
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4969..4985)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4993..5009
                     /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(5017..5047)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5062..5083
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5371..5959)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6130..6990)
                     /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(6991..7095)
                     /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 ggaaaattta aaaggttgca
      781 gtgtgtgctg taaatcttct tggaatcaac tgcaggacct gtgccgacta gccaagcttt
      841 cttgtcctgc ccttggtgtt tctaagaaga atctgtatga ctttgaagtt gaatacctgt
      901 gtgattataa gaagatccgt gagcaggagt attacctggt taagtggcgt gggtatcccg
      961 actcagaaaa cacctgggag ccacggcaga atctaaaatg tatacgagtt cttaagcagt
     1021 tccacaagga cttagaaaga gagcttgtcc gacgacaccg ccggtcaaag ccacccaggc
     1081 atctggaccc aaacctagcc aattacctgg tgcagaaggc caagcagagg cgggcacttc
     1141 agcgttggga acaagagctc aatgccaagc gcagccacct ggggcggatc accgtggaga
     1201 atgaggtaga cctggatggc cctccaaggt cctttgtcta tatcaatgag tatcgagttg
     1261 gtgagggcat caccctcaac caggtagctg ttggctgtga gtgccaggac tgtctgttgg
     1321 cacccactgg aggctgttgc cctggagcat ccctgcacaa gtttgcctac aatgaccaag
     1381 gccaggtgcg actgaaagct gggcagccca tctacgagtg caactcccgc tgttgctgtg
     1441 gctatgactg cccaaaccgt gtagtccaga aaggcatccg ctacgatctc tgcatcttcc
     1501 gcactaatga tggccgaggc tggggtgtcc gcacgctgga aaagatccga aaaaatagct
     1561 ttgttatgga gtatgtggga gagattatta cctcagagga ggcagagcgg aggggccaga
     1621 tctacgaccg ccagggcgcc acctacctct ttgacctgga ctacgtggaa gacgtatata
     1681 ccgtggatgc cgcttattat ggcaacatct ctcattttgt caaccatagt tgtgatccca
     1741 acctgcaggt gtacaacgta ttcatagaca accttgatga gcgactaccc cgcatcgcat
     1801 tctttgccac aagaaccatc tgggcgggcg aggagctcac ctttgattac aacatgcaag
     1861 tggaccccgt ggacatggag agtacccgaa tggactccaa ctttggcctg gctgggctcc
     1921 ccggctcccc caagaaacga gtccgtattg aatgcaaatg tgggacaacg gcttgccgaa
     1981 aatacctctt cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2041 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2101 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2161 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2221 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2281 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2341 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2401 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2461 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2521 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2581 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2641 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2701 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2761 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2821 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2881 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2941 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3001 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3061 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3121 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3181 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3241 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3301 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3361 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3421 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3661 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3781 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3841 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3901 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3961 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4021 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4081 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4141 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4201 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4261 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4321 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4381 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4441 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4501 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4561 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4621 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4681 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4741 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4801 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4861 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4921 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5101 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6001 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6061 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6121 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6181 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6241 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6301 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6361 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6421 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6481 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6541 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6601 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6661 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6721 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6781 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6841 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6901 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6961 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7021 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7081 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7141 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7201 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7261 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
