LOCUS       V019068                 7847 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019068
VERSION     V019068
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..2527
                     /label="VGF(NM_003378)"
                     /note="VGF(NM_003378)"
                     /gene="VGF"
     CDS             2534..2599
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2623..3211
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3277..3325
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3527..3955
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3969..4298
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4365..5159
                     /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    5335..5456
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5505..5521)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5529..5545
                     /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(5553..5583)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5598..5619
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5907..6495)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6666..7526)
                     /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(7527..7631)
                     /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 ccatgaaagc cctcagattg tcggcttccg ccctcttctg
      721 ccttctgctg atcaacgggt taggggcagc accccctggt cgccctgagg cgcagcctcc
      781 tcctctcagc tctgagcata aagagccggt agccggggac gcagtgcccg ggccaaagga
      841 tggcagcgcc ccagaggtcc gaggcgctcg gaattccgag ccgcaggacg agggagagct
      901 tttccagggc gtggatcccc gggcgctggc cgcggtgctg ctgcaggcac tcgaccgtcc
      961 cgcctcaccc ccggcaccaa gcggctccca gcaggggccg gaggaagaag cagctgaagc
     1021 tctgctgacc gagaccgtgc gcagccagac ccacagcctc ccggcgccgg agagcccgga
     1081 gcccgcggct ccgcctcgcc ctcagactcc ggagaatggg cccgaggcga gcgatccctc
     1141 cgaggagctc gaggcgctag cgtccctgct ccaggaactg cgagatttca gtccaagtag
     1201 cgccaagcgc cagcaggaga cggcggcagc agagacggaa acccgcacgc acacgctgac
     1261 ccgagtgaat ctggagagcc cggggccaga gcgcgtatgg cgcgcttcct ggggagagtt
     1321 ccaggcgcgt gtcccggagc gcgcgcccct gccgcccccg gccccctctc aattccaggc
     1381 gcgtatgccc gacagcgggc cccttcccga aacccacaag ttcggggaag gagtgtcctc
     1441 ccccaaaaca cacctaggcg aggcattggc acccctgtcc aaggcgtacc aaggcgtggc
     1501 cgccccgttc cccaaggcgc gccggccgga gagcgcactc ctgggcggct ccgaggcggg
     1561 cgagcgcctt ctccagcaag ggctggcgca ggtggaggcc gggcggcggc aggcggaggc
     1621 cacgcggcag gccgcggcgc aggaagagcg gctggccgac ctcgcctcgg acctgctgct
     1681 ccagtatttg ctgcagggcg gggcccggca gcgcggcctc gggggtcggg ggctgcagga
     1741 ggcggcggag gagcgagaga gtgcaaggga ggaggaggag gcggagcagg agagacgcgg
     1801 cggggaggag agggtggggg aagaggatga ggaggcggcc gaggcggagg cagaggcgga
     1861 ggaggcggag agggcgcggc agaacgcgct cctgttcgcg gaggaggagg acggggaagc
     1921 cggcgccgag gacaagcgct cccaggagga gacgccgggc caccggcgga aggaggccga
     1981 ggggacagag gagggcgggg aggaggagga cgacgaggag atggatccgc agacgatcga
     2041 cagcctcatt gagctgtcca ccaaactcca cctgccagcg gacgacgtgg tcagcatcat
     2101 cgaggaggtg gaggagaagc ggaagcggaa gaagaacgcc cctcccgagc ccgtgccgcc
     2161 cccccgtgcc gcccccgccc ccacccacgt ccgctccccg cagcccccgc cccccgcccc
     2221 cgctcccgca cgagacgagc tgccggactg gaacgaggtg ctcccgccct gggatcggga
     2281 ggaggacgag gtgtacccgc cagggccgta ccaccctttc cccaactaca tccggccgcg
     2341 gacactgcag ccgccctcgg ccttgcgccg ccgccactac caccacgcct tgccgccttc
     2401 gcgccactat cccggccggg aggcccaggc gcggcgcgcg caggaggagg cggaggcgga
     2461 ggagcgccgg ctgcaggagc aggaggagct ggagaattac atcgagcacg tgctgctccg
     2521 gcgcccgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2581 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     2641 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2701 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2761 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2821 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2881 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2941 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3001 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     3061 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3121 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3181 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     3241 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3301 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3361 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3421 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3481 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     3541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3661 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     3721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     3901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4141 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4201 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4261 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4321 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4381 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4441 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4501 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4561 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4621 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4681 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4741 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4801 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4861 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4921 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4981 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5041 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5101 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5161 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5221 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5281 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5341 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5401 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5461 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5521 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5581 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5641 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5701 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5761 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5821 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5881 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5941 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6001 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6061 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6121 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6181 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6241 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6301 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6361 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6421 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6481 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6541 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6601 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6661 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6721 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6781 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6841 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6901 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6961 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7021 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7081 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7141 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7201 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7261 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7321 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7381 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7441 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7501 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7561 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7621 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7681 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7741 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7801 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
