LOCUS       V035884                 8656 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035884
VERSION     V035884
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             706..3315
                     /label="WWP2(NM_001270454)"
                     /note="WWP2(NM_001270454)"
                     /gene="WWP2"
     CDS             3322..3348
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3352..3378
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3382..3408
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3432..4020
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4086..4134
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4336..4764
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4778..5107
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5174..5968
                     /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    6144..6265
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6314..6330)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6338..6354
                     /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(6362..6392)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6407..6428
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6716..7304)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7475..8335)
                     /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(8336..8440)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc atctgccagc
      721 tctagccggg caggagtggc cctgcctttt gagaagtctc agctcacttt gaaagtggtg
      781 tccgcaaagc ccaaggtgca taatcgtcaa cctcgaatta actcctacgt ggaggtggcg
      841 gtggatggac tccccagtga gaccaagaag actgggaagc gcattgggag ctctgagctt
      901 ctctggaatg agatcatcat tttgaatgtc acggcacaga gtcatttaga tttaaaggtc
      961 tggagctgcc ataccttgag aaatgaactg ctaggcaccg catctgtcaa cctctccaac
     1021 gtcttgaaga acaatggggg caaaatggag aacatgcagc tgaccctgaa cctgcagacg
     1081 gagaacaaag gcagcgttgt ctcaggcgga gagctgacaa ttttcctgga cgggccaact
     1141 gttgatctgg gaaatgtgcc taatggcagt gccctgacag atggatcaca gctgccttcg
     1201 agagactcca gtggaacagc agtagctcca gagaaccggc accagccccc cagcacaaac
     1261 tgctttggtg gaagatcccg gacgcacaga cattcgggtg cttcagccag aacaacccca
     1321 gcaaccggcg agcaaagccc cggtgctcgg agccggcacc gccagcccgt caagaactca
     1381 ggccacagtg gcttggccaa tggcacagtg aatgatgaac ccacaacagc cactgatccc
     1441 gaagaacctt ccgttgttgg tgtgacgtcc ccacctgctg cacccttgag tgtgaccccg
     1501 aatcccaaca cgacttctct ccctgcccca gccacaccgg ctgaaggaga ggaacccagc
     1561 acttcgggta cacagcagct cccagcggct gcccaggccc ccgacgctct gcctgctgga
     1621 tgggaacagc gagagctgcc caacggacgt gtctattatg ttgaccacaa taccaagacc
     1681 accacctggg agcggcccct tcctccaggc tgggaaaaac gcacagatcc ccgaggcagg
     1741 ttttactatg tggatcacaa tactcggacc accacctggc agcgtccgac cgcggagtac
     1801 gtgcgcaact atgagcagtg gcagtcgcag cggaatcagc tccagggggc catgcagcac
     1861 ttcagccaaa gattcctcta ccagtcttcg agtgcttcga ctgaccatga tcccctgggc
     1921 cccctccctc ctggctggga gaagagacag gacaatggac gggtgtatta cgtgaaccat
     1981 aacactcgca cgacccagtg ggaggatccc cggacccagg ggatgatcca ggaaccagct
     2041 ctgcccccag gatgggagat gaaatacacc agcgaggggg tgcgatactt tgtggaccac
     2101 aatacccgca ccaccacctt taaggatcct cgcccggggt ttgagtcggg gacgaagcaa
     2161 ggttcccctg gtgcttatga ccgcagtttt cggtggaagt atcaccagtt ccgtttcctc
     2221 tgccattcaa atgccctacc tagccacgtg aagatcagcg tttccaggca gacgcttttc
     2281 gaagattcct tccaacagat catgaacatg aaaccctatg acctgcgccg ccggctctac
     2341 atcatcatgc gtggcgagga gggcctggac tatgggggca tcgccagaga gtggtttttc
     2401 ctcctgtctc atgaggtgct caaccctatg tattgtttat ttgaatatgc cggaaagaac
     2461 aattactgcc tgcagatcaa ccccgcctcc tccatcaacc cggaccacct cacctacttt
     2521 cgctttatag gcagattcat cgccatggcg ctgtaccatg gaaagttcat cgacacgggc
     2581 ttcaccctcc ctttctacaa gcggatgctc aataagagac caaccctgaa agacctggag
     2641 tccattgacc ctgagttcta caactccatt gtctggatca aagagaacaa cctggaagaa
     2701 tgtggcctgg agctgtactt catccaggac atggagatac tgggcaaggt gacgacccac
     2761 gagctgaagg agggcggcga gagcatccgg gtcacagagg agaacaagga agagtacatc
     2821 atgctgctga ctgactggcg tttcacccga ggcgtggaag agcagaccaa agccttcctg
     2881 gatggcttca acgaggtggc cccgctggag tggctgcgct actttgacga gaaagagctg
     2941 gagctgatgc tgtgcggcat gcaggagata gacatgagcg actggcagaa gagcaccatc
     3001 taccggcact acaccaagaa cagcaagcag atccagtggt tctggcaggt ggtgaaggag
     3061 atggacaacg agaagaggat ccggctgctg cagtttgtca ccggtacctg ccgcctgccc
     3121 gtcgggggat ttgccgaact catcggtagc aacggaccac agaagttttg cattgacaaa
     3181 gttggcaagg aaacctggct gcccagaagc cacacctgct tcaaccgtct ggatcttcca
     3241 ccctacaaga gctacgaaca gctgagagag aagctgctgt atgccattga ggagaccgag
     3301 ggctttggac aggagctcga gtacccatac gacgtacctg attatgcaac ttatccttat
     3361 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gtgagggccc
     3421 gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3481 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3541 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3601 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3661 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3721 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3781 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     3841 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3901 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3961 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4021 ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4081 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
     4141 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4201 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4261 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
     4321 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt
     4501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4681 attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga
     4741 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4921 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4981 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5041 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5101 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     5161 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5341 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5701 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     6001 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6061 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6121 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6241 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6901 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7141 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8461 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     8521 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     8581 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     8641 gaagaatctg cttagg
//
