LOCUS       V019897                 7313 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019897
VERSION     V019897
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..1993
                     /label="E2F1(NM_005225)"
                     /note="E2F1(NM_005225)"
                     /gene="E2F1"
     CDS             2000..2065
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2089..2677
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2743..2791
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2993..3421
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3435..3764
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3831..4625
                     /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    4801..4922
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4971..4987)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4995..5011
                     /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(5019..5049)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5064..5085
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5373..5961)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6132..6992)
                     /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(6993..7097)
                     /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 ccatggcctt ggccggggcc cctgcgggcg gcccatgcgc
      721 gccggcgctg gaggccctgc tcggggccgg cgcgctgcgg ctgctcgact cctcgcagat
      781 cgtcatcatc tccgccgcgc aggacgccag cgccccgccg gctcccaccg gccccgcggc
      841 gcccgccgcc ggcccctgcg accctgacct gctgctcttc gccacaccgc aggcgccccg
      901 gcccacaccc agtgcgccgc ggcccgcgct cggccgcccg ccggtgaagc ggaggctgga
      961 cctggaaact gaccatcagt acctggccga gagcagtggg ccagctcggg gcagaggccg
     1021 ccatccagga aaaggtgtga aatccccggg ggagaagtca cgctatgaga cctcactgaa
     1081 tctgaccacc aagcgcttcc tggagctgct gagccactcg gctgacggtg tcgtcgacct
     1141 gaactgggct gccgaggtgc tgaaggtgca gaagcggcgc atctatgaca tcaccaacgt
     1201 ccttgagggc atccagctca ttgccaagaa gtccaagaac cacatccagt ggctgggcag
     1261 ccacaccaca gtgggcgtcg gcggacggct tgaggggttg acccaggacc tccgacagct
     1321 gcaggagagc gagcagcagc tggaccacct gatgaatatc tgtactacgc agctgcgcct
     1381 gctctccgag gacactgaca gccagcgcct ggcctacgtg acgtgtcagg accttcgtag
     1441 cattgcagac cctgcagagc agatggttat ggtgatcaaa gcccctcctg agacccagct
     1501 ccaagccgtg gactcttcgg agaactttca gatctccctt aagagcaaac aaggcccgat
     1561 cgatgttttc ctgtgccctg aggagaccgt aggtgggatc agccctggga agaccccatc
     1621 ccaggaggtc acttctgagg aggagaacag ggccactgac tctgccacca tagtgtcacc
     1681 accaccatca tctcccccct catccctcac cacagatccc agccagtctc tactcagcct
     1741 ggagcaagaa ccgctgttgt cccggatggg cagcctgcgg gctcccgtgg acgaggaccg
     1801 cctgtccccg ctggtggcgg ccgactcgct cctggagcat gtgcgggagg acttctccgg
     1861 cctcctccct gaggagttca tcagcctttc cccaccccac gaggccctcg actaccactt
     1921 cggcctcgag gagggcgagg gcatcagaga cctcttcgac tgtgactttg gggacctcac
     1981 ccccctggat ttcctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     2041 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     2101 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2161 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2221 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2281 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2341 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2401 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2461 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     2521 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2581 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2641 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     2701 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     2761 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     2821 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     2881 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     2941 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     3001 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3061 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3121 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     3181 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3301 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3361 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3421 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3541 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3601 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3661 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3721 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3781 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3841 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3901 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3961 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4021 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4081 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4141 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4201 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4261 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4321 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4381 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4441 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4501 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4561 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4621 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     4681 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4741 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4801 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4861 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4921 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4981 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5041 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5161 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7141 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     7201 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     7261 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
