LOCUS       V022812                 8188 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022812
VERSION     V022812
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..2868
                     /label="WDCP(NM_025203)"
                     /note="WDCP(NM_025203)"
                     /gene="WDCP"
     CDS             2875..2940
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2964..3552
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3618..3666
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3868..4296
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4310..4639
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4706..5500
                     /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    5676..5797
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5846..5862)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5870..5886
                     /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(5894..5924)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5939..5960
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6248..6836)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7007..7867)
                     /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(7868..7972)
                     /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 ccaccatgga gttgggaaaa
      721 ggaaaactac tcaggactgg actgaatgcg ttgcatcaag cagtgcatcc gatccatggc
      781 cttgcctgga ccgatgggaa tcaagttgtc ctaactgatt tgcggcttca cagtggagag
      841 gtcaagtttg gggactccaa agtcattgga cagtttgaat gtgtctgtgg gttgtcctgg
      901 gccccacctg ttgcagatga tacacctgtt ctactcgctg tccagcatga gaagcatgtc
      961 actgtgtggc agctgtgtcc cagccctatg gagtcaagca aatggctgac gtctcagact
     1021 tgtgagatta gaggatcact acctatcctt ccccagggct gtgtgtggca cccaaaatgt
     1081 gctattctga ctgtgttgac tgctcaggat gtctccattt tccctaatgt tcactctgat
     1141 gattcccagg taaaggcaga catcaacacc cagggccgca ttcactgtgc atgttggacc
     1201 caggatggcc tgaggctggt ggtggcagta ggcagcagcc tgcattctta tatttgggac
     1261 agcgctcaga agactcttca caggtgctcc tcctgcctgg tgtttgatgt ggacagccac
     1321 gtctgctcca tcacagcaac tgtggactca caggttgcta tagctactga gcttccattg
     1381 gataagatct gtggcttaaa tgcatctgaa acctttaata tcccacctaa cagtaaagac
     1441 atgactccgt atgctttacc agttattggt gaagtacgct ctatggataa agaggcaact
     1501 gattctgaaa caaattctga agtatcagtt tcttcttcct atttagaacc tctggatcta
     1561 actcacatac atttcaatca acataagtct gagggtaatt ctcttatttg tctaagaaaa
     1621 aaggactact tgacaggaac tggccaagat tcttcacatt tggtccttgt gacctttaag
     1681 aaggcagtta ccatgacgag aaaagtcact attccaggca ttctggttcc tgatctgata
     1741 gcatttaatc ttaaagccca cgtagtggca gtggcttcca acacttgtaa tataattttg
     1801 atctactctg tcattccatc ttcagtccca aacatccagc aaattcgatt agagaacact
     1861 gaaagaccaa aagggatatg tttcttgaca gaccaactat tactaatttt ggtaggaaaa
     1921 caaaaactca ctgatacaac atttcttcct tcttcaaagt ctgatcagta tgccattagc
     1981 ttgattgtta gagaaataat gttggaagaa gaaccttcaa taacatcagg tgaaagccag
     2041 actacctact ctactttcag tgctccgtta aataaagcaa atagaaaaaa gttaattgaa
     2101 agtctttccc cagatttttg tcaccaaaac aaagggctgt tgctgacagt taataccagt
     2161 agtcagaatg gaaggcctgg aagaacactt attaaagaaa tccagagtcc tctgtctagt
     2221 atctgtgatg gctccatagc tctagatgct gagcctgtta cccagccagc atcgctgccc
     2281 agacacagca gcacaccaga ccacaccagc acactggagc ctcctcgttt gcctcaaaga
     2341 aagaacttac aaagtgaaaa ggaaacttat cagctgtcta aggaagtgga aattttatct
     2401 aggaacctgg ttgaaatgca acggtgtctt tctgaactta caaaccgtct gcataatggg
     2461 aagaaatcct cttcagtgta tccactctct caagatcttc cttatgttca catcatttac
     2521 cagaaacctt attatctagg tcctgttgtt gaaaaaagag cggtgcttct ctgtgatggt
     2581 aaactaaggc tcagtacagt tcagcagact tttggccttt ctctcattga aatgctacat
     2641 gattcccact ggattcttct ctctgctgac agtgagggct ttatcccgtt aaccttcaca
     2701 gccacacagg aaataatcat aagagatggc agcctgtcca ggtcagatgt cttcagagac
     2761 tctttttctc acagtccagg tgctgtttct tctcttaaag tctttacagg ccttgctgcc
     2821 cccagtttag ataccactgg ctgttgtaac catgtagatg gcatggctct cgaggactac
     2881 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     2941 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     3001 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3061 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3121 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3181 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3241 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3301 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     3361 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3421 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3481 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3541 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     3601 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     3661 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     3721 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     3781 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     3841 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3901 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3961 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4021 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4081 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4141 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4201 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     4261 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4321 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4381 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4441 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4501 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4561 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4621 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4681 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4741 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4801 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4861 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4921 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4981 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5041 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5101 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5161 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5221 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5281 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5341 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5401 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5461 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     5521 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5581 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5641 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5701 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5761 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5821 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5881 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5941 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6001 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6061 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6421 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     6481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6661 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7081 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7141 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7201 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7261 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7321 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7381 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7441 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7501 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7561 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7621 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7681 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7741 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7801 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7861 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7921 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7981 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     8041 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     8101 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     8161 cgacaattgc atgaagaatc tgcttagg
//
