LOCUS       V018557                 8060 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018557
VERSION     V018557
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             941..3550
                     /label="WWP2(NM_007014)"
                     /note="WWP2(NM_007014)"
                     /gene="WWP2"
     CDS             3557..3622
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3660..3884
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3930..4358
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4372..4701
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4768..5562
                     /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    5736..5857
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5906..5922)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5930..5946
                     /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(5954..5984)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5999..6020
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6308..6893)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7064..7924)
                     /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(7925..8029)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcatctg ccagctctag
      961 ccgggcagga gtggccctgc cttttgagaa gtctcagctc actttgaaag tggtgtccgc
     1021 aaagcccaag gtgcataatc gtcaacctcg aattaactcc tacgtggagg tggcggtgga
     1081 tggactcccc agtgagacca agaagactgg gaagcgcatt gggagctctg agcttctctg
     1141 gaatgagatc atcattttga atgtcacggc acagagtcat ttagatttaa aggtctggag
     1201 ctgccatacc ttgagaaatg aactgctagg caccgcatct gtcaacctct ccaacgtctt
     1261 gaagaacaat gggggcaaaa tggagaacat gcagctgacc ctgaacctgc agacggagaa
     1321 caaaggcagc gttgtctcag gcggagagct gacaattttc ctggacgggc caactgttga
     1381 tctgggaaat gtgcctaatg gcagtgccct gacagatgga tcacagctgc cttcgagaga
     1441 ctccagtgga acagcagtag ctccagagaa ccggcaccag ccccccagca caaactgctt
     1501 tggtggaaga tcccggacgc acagacattc gggtgcttca gccagaacaa ccccagcaac
     1561 cggcgagcaa agccccggtg ctcggagccg gcaccgccag cccgtcaaga actcaggcca
     1621 cagtggcttg gccaatggca cagtgaatga tgaacccaca acagccactg atcccgaaga
     1681 accttccgtt gttggtgtga cgtccccacc tgctgcaccc ttgagtgtga ccccgaatcc
     1741 caacacgact tctctccctg ccccagccac accggctgaa ggagaggaac ccagcacttc
     1801 gggtacacag cagctcccag cggctgccca ggcccccgac gctctgcctg ctggatggga
     1861 acagcgagag ctgcccaacg gacgtgtcta ttatgttgac cacaatacca agaccaccac
     1921 ctgggagcgg ccccttcctc caggctggga aaaacgcaca gatccccgag gcaggtttta
     1981 ctatgtggat cacaatactc ggaccaccac ctggcagcgt ccgaccgcgg agtacgtgcg
     2041 caactatgag cagtggcagt cgcagcggaa tcagctccag ggggccatgc agcacttcag
     2101 ccaaagattc ctctaccagt cttcgagtgc ttcgactgac catgatcccc tgggccccct
     2161 ccctcctggc tgggagaaga gacaggacaa tggacgggtg tattacgtga accataacac
     2221 tcgcacgacc cagtgggagg atccccggac ccaggggatg atccaggaac cagctctgcc
     2281 cccaggatgg gagatgaaat acaccagcga gggggtgcga tactttgtgg accacaatac
     2341 ccgcaccacc acctttaagg atcctcgccc ggggtttgag tcggggacga agcaaggttc
     2401 ccctggtgct tatgaccgca gttttcggtg gaagtatcac cagttccgtt tcctctgcca
     2461 ttcaaatgcc ctacctagcc acgtgaagat cagcgtttcc aggcagacgc ttttcgaaga
     2521 ttccttccaa cagatcatga acatgaaacc ctatgacctg cgccgccggc tctacatcat
     2581 catgcgtggc gaggagggcc tggactatgg gggcatcgcc agagagtggt ttttcctcct
     2641 gtctcatgag gtgctcaacc ctatgtattg tttatttgaa tatgccggaa agaacaatta
     2701 ctgcctgcag atcaaccccg cctcctccat caacccggac cacctcacct actttcgctt
     2761 tataggcaga ttcatcgcca tggcgctgta ccatggaaag ttcatcgaca cgggcttcac
     2821 cctccctttc tacaagcgga tgctcaataa gagaccaacc ctgaaagacc tggagtccat
     2881 tgaccctgag ttctacaact ccattgtctg gatcaaagag aacaacctgg aagaatgtgg
     2941 cctggagctg tacttcatcc aggacatgga gatactgggc aaggtgacga cccacgagct
     3001 gaaggagggc ggcgagagca tccgggtcac agaggagaac aaggaagagt acatcatgct
     3061 gctgactgac tggcgtttca cccgaggcgt ggaagagcag accaaagcct tcctggatgg
     3121 cttcaacgag gtggccccgc tggagtggct gcgctacttt gacgagaaag agctggagct
     3181 gatgctgtgc ggcatgcagg agatagacat gagcgactgg cagaagagca ccatctaccg
     3241 gcactacacc aagaacagca agcagatcca gtggttctgg caggtggtga aggagatgga
     3301 caacgagaag aggatccggc tgctgcagtt tgtcaccggt acctgccgcc tgcccgtcgg
     3361 gggatttgcc gaactcatcg gtagcaacgg accacagaag ttttgcattg acaaagttgg
     3421 caaggaaacc tggctgccca gaagccacac ctgcttcaac cgtctggatc ttccacccta
     3481 caagagctac gaacagctga gagagaagct gctgtatgcc attgaggaga ccgagggctt
     3541 tggacaggag ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3601 ctacaaggat gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     3661 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3721 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3781 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3841 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3901 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4381 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4441 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4501 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4621 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4681 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4741 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4801 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4861 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4921 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4981 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5041 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5101 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5161 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5221 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5281 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5341 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5401 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5461 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5521 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5581 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5641 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5701 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5881 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6061 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6961 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7021 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7081 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7141 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7201 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7261 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7321 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7381 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7441 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7501 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7561 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7621 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7681 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7741 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7801 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7861 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7921 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8041 gaaaagtgcc acctgacgtc
//
