LOCUS       V027731                 8196 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027731
VERSION     V027731
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             919..3684
                     /label="WWP1(NM_007013)"
                     /note="WWP1(NM_007013)"
                     /gene="WWP1"
     CDS             3691..3717
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        complement(3748..3766)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    3792..4016
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4062..4490
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4504..4834
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4901..5695
                     /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    5869..5990
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6039..6055)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6063..6079
                     /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(6087..6117)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6132..6153
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6441..7029)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7200..8060)
                     /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(8061..8165)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccgccaccat ggccactgct tcaccaaggt ctgatactag taataaccac
      961 agtggaaggt tgcagttaca ggtaactgtt tctagtgcca aacttaaaag aaaaaagaac
     1021 tggttcggaa cagcaatata tacagaagta gttgtagatg gagaaattac gaaaacagca
     1081 aaatccagta gttcttctaa tccaaaatgg gatgaacagc taactgtaaa tgttacgcca
     1141 cagactacat tggaatttca agtttggagc catcgcactt taaaagcaga tgctttatta
     1201 ggaaaagcaa cgatagattt gaaacaagct ctgttgatac acaatagaaa attggaaaga
     1261 gtgaaagaac aattaaaact ttccttggaa aacaagaatg gcatagcaca aactggtgaa
     1321 ttgacagttg tgcttgatgg attggtgatt gagcaagaaa atataacaaa ctgcagctca
     1381 tctccaacca tagaaataca ggaaaatggt gatgccttac atgaaaatgg agagccttca
     1441 gcaaggacaa ctgccaggtt ggctgttgaa ggcacgaatg gaatagataa tcatgtacct
     1501 acaagcactc tagtccaaaa ctcatgctgc tcgtatgtag ttaatggaga caacacacct
     1561 tcatctccgt ctcaggttgc tgccagaccc aaaaatacac cagctccaaa accactcgca
     1621 tctgagcctg ccgatgacac tgttaatgga gaatcatcct catttgcacc aactgataat
     1681 gcgtctgtca cgggtactcc agtagtgtct gaagaaaatg ccttgtctcc aaattgcact
     1741 agtactactg ttgaagatcc tccagttcaa gaaatactga cttcctcaga aaacaatgaa
     1801 tgtattcctt ctaccagtgc agaattggaa tctgaagcta gaagtatatt agagcctgac
     1861 acctctaatt ctagaagtag ttctgctttt gaagcagcca aatcaagaca gccagatggg
     1921 tgtatggatc ctgtacggca gcagtctggg aatgccaaca cagaaacctt gccatcaggg
     1981 tgggaacaaa gaaaagatcc tcatggtaga acctattatg tggatcataa tactcgaact
     2041 accacatggg agagaccaca acctttacct ccaggttggg aaagaagagt tgatgatcgt
     2101 agaagagttt attatgtgga tcataacacc agaacaacaa cgtggcagcg gcctaccatg
     2161 gaatctgtcc gaaattttga acagtggcaa tctcagcgga accaattgca gggagctatg
     2221 caacagttta accaacgata cctctattcg gcttcaatgt tagctgcaga aaatgaccct
     2281 tatggacctt tgccaccagg ctgggaaaaa agagtggatt caacagacag ggtttacttt
     2341 gtgaatcata acacaaaaac aacccagtgg gaagatccaa gaactcaagg cttacagaat
     2401 gaagaacccc tgccagaagg ctgggaaatt agatatactc gtgaaggtgt aaggtacttt
     2461 gttgatcata acacaagaac aacaacattc aaagatcctc gcaatgggaa gtcatctgta
     2521 actaaaggtg gtccacaaat tgcttatgaa cgcggcttta ggtggaagct tgctcacttc
     2581 cgttatttgt gccagtctaa tgcactacct agtcatgtaa agatcaatgt gtcccggcag
     2641 acattgtttg aagattcctt ccaacagatt atggcattaa aaccctatga cttgaggagg
     2701 cgcttatatg taatatttag aggagaagaa ggacttgatt atggtggcct agcgagagaa
     2761 tggtttttct tgctttcaca tgaagttttg aacccaatgt attgcttatt tgagtatgcg
     2821 ggcaagaaca actattgtct gcagataaat ccagcatcaa ccattaatcc agaccatctt
     2881 tcatacttct gtttcattgg tcgttttatt gccatggcac tatttcatgg aaagtttatc
     2941 gatactggtt tctctttacc attctacaag cgtatgttaa gtaaaaaact tactattaag
     3001 gatttggaat ctattgatac tgaattttat aactccctta tctggataag agataacaac
     3061 attgaagaat gtggcttaga aatgtacttt tctgttgaca tggagatttt gggaaaagtt
     3121 acttcacatg acctgaagtt gggaggttcc aatattctgg tgactgagga gaacaaagat
     3181 gaatatattg gtttaatgac agaatggcgt ttttctcgag gagtacaaga acagaccaaa
     3241 gctttccttg atggttttaa tgaagttgtt cctcttcagt ggctacagta cttcgatgaa
     3301 aaagaattag aggttatgtt gtgtggcatg caggaggttg acttggcaga ttggcagaga
     3361 aatactgttt atcgacatta tacaagaaac agcaagcaaa tcatttggtt ttggcagttt
     3421 gtgaaagaga cagacaatga agtaagaatg cgactattgc agttcgtcac tggaacctgc
     3481 cgtttacctc taggaggatt tgctgagctc atgggaagta atgggcctca aaagttttgc
     3541 attgaaaaag ttggcaaaga cacttggtta ccaagaagcc atacatgttt taatcgcttg
     3601 gatctaccac catataagag ttatgaacaa ctaaaggaaa aacttctttt tgcaatagaa
     3661 gagacagagg gatttggaca agaagaattc tacccatacg acgtaccaga ttacgcttga
     3721 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     3781 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3841 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3901 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3961 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4021 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4081 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4141 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4201 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4261 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4321 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4381 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4441 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4501 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     4561 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4621 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4681 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4741 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4801 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4861 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4921 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4981 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5041 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5101 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5161 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5221 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5281 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5341 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5401 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5461 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5521 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5581 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5641 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5701 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5761 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5821 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6001 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6601 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6841 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8161 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
