LOCUS       V024991                 8841 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024991
VERSION     V024991
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3521
                     /label="WWP1(NM_007013)"
                     /note="WWP1(NM_007013)"
                     /gene="WWP1"
     CDS             3528..3593
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3617..4205
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4271..4319
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4521..4949
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4963..5292
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5359..6153
                     /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    6329..6450
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6499..6515)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6523..6539
                     /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(6547..6577)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6592..6613
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6901..7489)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7660..8520)
                     /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(8521..8625)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cactgcttca ccaaggtctg
      781 atactagtaa taaccacagt ggaaggttgc agttacaggt aactgtttct agtgccaaac
      841 ttaaaagaaa aaagaactgg ttcggaacag caatatatac agaagtagtt gtagatggag
      901 aaattacgaa aacagcaaaa tccagtagtt cttctaatcc aaaatgggat gaacagctaa
      961 ctgtaaatgt tacgccacag actacattgg aatttcaagt ttggagccat cgcactttaa
     1021 aagcagatgc tttattagga aaagcaacga tagatttgaa acaagctctg ttgatacaca
     1081 atagaaaatt ggaaagagtg aaagaacaat taaaactttc cttggaaaac aagaatggca
     1141 tagcacaaac tggtgaattg acagttgtgc ttgatggatt ggtgattgag caagaaaata
     1201 taacaaactg cagctcatct ccaaccatag aaatacagga aaatggtgat gccttacatg
     1261 aaaatggaga gccttcagca aggacaactg ccaggttggc tgttgaaggc acgaatggaa
     1321 tagataatca tgtacctaca agcactctag tccaaaactc atgctgctcg tatgtagtta
     1381 atggagacaa cacaccttca tctccgtctc aggttgctgc cagacccaaa aatacaccag
     1441 ctccaaaacc actcgcatct gagcctgccg atgacactgt taatggagaa tcatcctcat
     1501 ttgcaccaac tgataatgcg tctgtcacgg gtactccagt agtgtctgaa gaaaatgcct
     1561 tgtctccaaa ttgcactagt actactgttg aagatcctcc agttcaagaa atactgactt
     1621 cctcagaaaa caatgaatgt attccttcta ccagtgcaga attggaatct gaagctagaa
     1681 gtatattaga gcctgacacc tctaattcta gaagtagttc tgcttttgaa gcagccaaat
     1741 caagacagcc agatgggtgt atggatcctg tacggcagca gtctgggaat gccaacacag
     1801 aaaccttgcc atcagggtgg gaacaaagaa aagatcctca tggtagaacc tattatgtgg
     1861 atcataatac tcgaactacc acatgggaga gaccacaacc tttacctcca ggttgggaaa
     1921 gaagagttga tgatcgtaga agagtttatt atgtggatca taacaccaga acaacaacgt
     1981 ggcagcggcc taccatggaa tctgtccgaa attttgaaca gtggcaatct cagcggaacc
     2041 aattgcaggg agctatgcaa cagtttaacc aacgatacct ctattcggct tcaatgttag
     2101 ctgcagaaaa tgacccttat ggacctttgc caccaggctg ggaaaaaaga gtggattcaa
     2161 cagacagggt ttactttgtg aatcataaca caaaaacaac ccagtgggaa gatccaagaa
     2221 ctcaaggctt acagaatgaa gaacccctgc cagaaggctg ggaaattaga tatactcgtg
     2281 aaggtgtaag gtactttgtt gatcataaca caagaacaac aacattcaaa gatcctcgca
     2341 atgggaagtc atctgtaact aaaggtggtc cacaaattgc ttatgaacgc ggctttaggt
     2401 ggaagcttgc tcacttccgt tatttgtgcc agtctaatgc actacctagt catgtaaaga
     2461 tcaatgtgtc ccggcagaca ttgtttgaag attccttcca acagattatg gcattaaaac
     2521 cctatgactt gaggaggcgc ttatatgtaa tatttagagg agaagaagga cttgattatg
     2581 gtggcctagc gagagaatgg tttttcttgc tttcacatga agttttgaac ccaatgtatt
     2641 gcttatttga gtatgcgggc aagaacaact attgtctgca gataaatcca gcatcaacca
     2701 ttaatccaga ccatctttca tacttctgtt tcattggtcg ttttattgcc atggcactat
     2761 ttcatggaaa gtttatcgat actggtttct ctttaccatt ctacaagcgt atgttaagta
     2821 aaaaacttac tattaaggat ttggaatcta ttgatactga attttataac tcccttatct
     2881 ggataagaga taacaacatt gaagaatgtg gcttagaaat gtacttttct gttgacatgg
     2941 agattttggg aaaagttact tcacatgacc tgaagttggg aggttccaat attctggtga
     3001 ctgaggagaa caaagatgaa tatattggtt taatgacaga atggcgtttt tctcgaggag
     3061 tacaagaaca gaccaaagct ttccttgatg gttttaatga agttgttcct cttcagtggc
     3121 tacagtactt cgatgaaaaa gaattagagg ttatgttgtg tggcatgcag gaggttgact
     3181 tggcagattg gcagagaaat actgtttatc gacattatac aagaaacagc aagcaaatca
     3241 tttggttttg gcagtttgtg aaagagacag acaatgaagt aagaatgcga ctattgcagt
     3301 tcgtcactgg aacctgccgt ttacctctag gaggatttgc tgagctcatg ggaagtaatg
     3361 ggcctcaaaa gttttgcatt gaaaaagttg gcaaagacac ttggttacca agaagccata
     3421 catgttttaa tcgcttggat ctaccaccat ataagagtta tgaacaacta aaggaaaaac
     3481 ttctttttgc aatagaagag acagagggat ttggacaaga aggatccgac tacaaagacc
     3541 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     3601 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3661 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3721 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3781 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3841 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3901 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3961 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     4021 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4081 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4141 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4201 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4261 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     4321 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4381 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4441 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     4501 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     4681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4861 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     4921 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5221 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5281 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5341 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5401 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5461 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5521 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5581 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5641 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5701 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5761 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5821 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5881 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5941 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6001 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6061 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6121 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     6181 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6241 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6301 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6421 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6481 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6541 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6601 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6661 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7081 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7321 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8641 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     8701 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     8761 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     8821 tgcatgaaga atctgcttag g
//
