LOCUS       V020793                 7457 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020793
VERSION     V020793
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..2137
                     /label="Yap1(NM_001394328)"
                     /note="Yap1(NM_001394328)"
                     /gene="Yap1"
     CDS             2144..2209
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2233..2821
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2887..2935
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3137..3565
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3579..3908
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3975..4769
                     /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    4945..5066
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5115..5131)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5139..5155
                     /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(5163..5193)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5208..5229
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5517..6105)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6276..7136)
                     /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(7137..7241)
                     /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 ccatggagcc cgcgcaacag ccgccgcccc agccggcccc
      721 gcaaggcccc gcgccgccgt cggtgtctcc ggccgggacc cccgcggccc cgcccgcacc
      781 cccggcgggt caccaggtcg tgcacgtccg cggggactcg gagaccgacc tggaagcgct
      841 tttcaacgcc gtcatgaacc ccaagacggc caacgtgcca cagaccgtgc ccatgaggct
      901 tcgcaagctg cccgactcct tcttcaagcc gcctgagccc aagtcccact cgcgacaggc
      961 cagtaccgat gcgggcactg ctggagccct gactccccag cacgttcgag ctcactcgtc
     1021 tccagcctcc ctgcagctgg gggccgggac actcacggcc agtggtgttg tctctggccc
     1081 ggccgccacc cctgctgctc aacatctcag acagtcttcc tttgagatcc ctgatgatgt
     1141 accattgcca gcaggctggg agatggccaa gacctcttct ggtcagagat acttcttaaa
     1201 tcacaatgat cagacaacaa catggcagga cccccggaag gccatgctct cccaactgaa
     1261 cgttcctaca tctgccagcc cagcagtgcc ccagacgctg atgaactctg cctcagggcc
     1321 tcttcctgat ggatgggagc aagccatgac tcaggatgga gaagtttact acataaacca
     1381 taagaacaag accacatcct ggctggaccc aaggcttgac cctcgttttg ccatgaacca
     1441 gaggatcact cagagtgctc cagtgaagca gcccccaccc ttggctcccc agagcccaca
     1501 gggaggcgtc ctgggtggag gcagctcaaa ccagcagcag cagatacagc tgcagcagct
     1561 acagatggag aaggagaggc tgcgattgaa acagcaggag ttatttcggc aggcaatacg
     1621 gaatatcaat cccagcacag caaatgctcc aaaatgtcag gaattagctc tccgcagcca
     1681 gttgccctca ctggagcagg atggagggac tcagaatgca gtgtcttctc ccgggatgac
     1741 tcaggaattg aggacaatga caaccaatag ttcagatccc tttcttaaca gtggcaccta
     1801 tcactctaga gatgagagca cagacagtgg cctcagcatg agcagctaca gcatccctcg
     1861 gaccccagac gacttcctga acagcgtgga tgaaatggac acaggagaca ccatcagcca
     1921 gagcaccctg ccgtcccagc agagccgctt ccctgattac ctggaggccc tccctgggac
     1981 aaatgtggac cttggcacac tggaaggaga tgcaatgaac atagaagggg aggagctgat
     2041 gcccagcctt caggaagcgc tgagttccga aatcctggat gtggagtctg tgttggctgc
     2101 caccaagtta gataaagaaa gctttctcac gtggttactc gaggactaca aagaccatga
     2161 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2221 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2281 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2341 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2401 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2461 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2521 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2581 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     2641 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2701 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2761 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2821 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     2881 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     2941 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3001 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3061 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3121 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3181 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3241 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3301 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3361 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3421 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3481 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3541 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3661 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3781 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3841 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3901 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3961 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4021 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4081 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4141 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4201 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4261 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4321 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4381 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4441 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4501 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4561 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4621 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4681 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4741 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4801 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4861 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4921 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4981 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5041 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5101 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5701 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5941 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7261 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     7321 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7381 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     7441 tgaagaatct gcttagg
//
