LOCUS       V046982                 9768 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046982
VERSION     V046982
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..4448
                     /label="Kif4(NM_008446)"
                     /note="Kif4(NM_008446)"
                     /gene="Kif4"
     CDS             4455..4520
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4544..5132
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5198..5246
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5448..5876
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5890..6219
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6286..7080
                     /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    7256..7377
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7426..7442)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7450..7466
                     /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(7474..7504)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7519..7540
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7828..8416)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8587..9447)
                     /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(9448..9552)
                     /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 ccaccatgaa agaagaggtg aaggggattc
      781 ccgtaagagt ggcactgcgt tgtcgccccc tcgtctcgaa agagattaag gagggctgcc
      841 agacgtgcct ttcctttgtg cccggagagc ctcaggtggt ggttggtaat gataaatcat
      901 ttacctacga ttttgtgttt gacccctcta ctgaacagga agaggtcttt aatacagcag
      961 tagctccact cataaaaggc gtgtttaaag gatacaatgc aactgtcctg gcctatgggc
     1021 agactggatc tggaaaaacg tattcaatgg gaggtgcata cactgcagaa caagaacatg
     1081 actcagccat tggggttatt cccagggtaa tacaactgct cttcaaagaa attaataaaa
     1141 agagtgactt cgaatttact ctgaaagtgt cttacttgga gatctataat gaagaaattt
     1201 tggaccttct atgttcatct cgtgagaaag ctactcaaat aaatatccgg gaggacccta
     1261 aggaaggcat aaagattgtg ggactgacag agaagactgt gctggtggcc tcggacaccg
     1321 tttcctgtct agagcagggt aacaactcca ggactgtggc ctccacagct atgaactccc
     1381 agtcatctcg atctcatgcc atctttacaa tctccattga gcaaaggaag aagaatgaca
     1441 aaaatagcag ctttcgttcc aagctgcatc ttgtagacct cgctggatca gaaagacaga
     1501 agaaaaccaa ggctgaggga gatcgactga gagagggtat taatattaat cgaggccttc
     1561 tgtgcttggg gaatgtaatc agtgctcttg gagatgataa aaagggtaac tttgtgccct
     1621 acagagattc caagctgact cgactgcttc aagattctct aggaggcaat agccacactc
     1681 ttatgattgc ctgtgtgagt cctgctgatt ccaatctaga agaaacatta aatacccttc
     1741 gctatgctga cagagcaaga aaaatcaaga acaaacctat tattaatatt gatccccagg
     1801 cagctgaact aaatcatctt aaacaacagg tccaacagct gcagatcttg ctgctacaag
     1861 cccatggagg caccttgcct ggagatataa atgtggaacc atcagaaaat cttcagtccc
     1921 tgatggagaa gaatcagtct ctggtggaag agaatgaaaa attaagtcgt ggtctgagtg
     1981 aggcagctgg tcaaacagcc cagatgctgg agaggatcat tctgacagag caagcaaatg
     2041 agaaaatgaa tgccaagcta gaagaactca ggcggcatgc ggcctgtaaa gtggatcttc
     2101 aaaagctcgt ggagacattg gaagaccagg agttaaaaga aaacatagag ataatttgca
     2161 acctacaaca agtgattgcc cagctatctg atgaagctgc tgcttgcatg actgcaacca
     2221 ttgatactgc aggggaagca gacactcaag tgcaaagcag cccagatacc agcaggtctt
     2281 ctgatgtttt cagcactcag catgccctac gccaagctca gatgtccaag gagctgattg
     2341 agttgaataa agcacttgca ctgaaggaag ccctagctaa gaagatgact cagaatgaca
     2401 accaactcca gcccattcag ttccagtacc aggataatat taaaaatcta gaatcagaag
     2461 tcctcagtct acagagggaa aaagaagaac tggttcttga acttcaaaca gcaaagaagg
     2521 atgccaacca agccaagttg agtgagcgcc gtcgtaaacg tcttcaggaa ctggagggtc
     2581 aaatagctga tttgaaaaag aaactgcaag aacagtccaa acttctgaag ctgaaggaat
     2641 ccacagagca tactgtctcc aagctgaacc aggagattcg gatgatgaaa aaccagcgag
     2701 tccagttaat gcgtcaaatg aaagaggatg ctgagaagtt taggcaatgg aagcagcaaa
     2761 aagacaaaga agtaatccag ctaaaagaac gggatcgtaa aaggcaatat gagctgctca
     2821 aacttgaaag aaactttcag aagcagtcta atgtgctcag acgtaaaact gaggaggcag
     2881 cagctgccaa caaacgtctt aaggatgctc tgcaaaagca aaaggaggtt gcagaaaaac
     2941 gcaaagagac tcagagccgt gggatggaaa gcactgcagc tagaatgaag aattggcttg
     3001 gaaatgaaat tgaggttatg gtcagtactg aggaagccaa gcgacatctg aatggccttc
     3061 ttgaagagag aaagatcctg gctcaggatg tggcacaact caaagagaaa agagaatctg
     3121 gagagaaccc acctcttaaa ctccgaaggc gcacattttc ctacgatgaa atacatggtc
     3181 aagattcagg agcagaggat tctattgcca agcagattga aagcctagag accgaattgg
     3241 agctcaggag tgcgcagatt gcggacctac agcaaaagct gctggatgct gaaagtgaag
     3301 accggccgaa acagcgctgg gagagtatcg ccacaattct cgaagccaaa tgtgccataa
     3361 agtatttagt gggagagctc gtctcctcca aaatactggt gagcaaacta gaaagcagcc
     3421 tgaatcagag caaggccagc tgtattgacg tgcagaagat gctgtttgag gagcaaaatc
     3481 attttgctaa gatagagaca gagttaaaag aagagctggt caaagtggag caacagcacc
     3541 aagaaaaggt gttgtacctt ctcagccagc tgcagcaatc ccagatgaca gagaagcagc
     3601 tggaggagtc agttagtgaa aaggaacagc agctgctgag cacactaaaa tgtcaggagg
     3661 aagagcttag gaagatgcaa gaagtgtgtg agcaaaatca gcagcttctc caagaaaaca
     3721 gtgccatcaa gcagaaactg accctactcc aagtagccag caaacagaaa ccccatctca
     3781 caagaaatat cttccagtct ccagactctt cctttgaata tattccacct aagcccaaac
     3841 cttgccgcat taaggaaaaa tgcctagagc aaagcttcgc cgtcgagggg ctgcagtatt
     3901 attcagagcc ttctgtggcc gagcaggaca atgaggacag tgacgaccat gctgatgagg
     3961 agtggatccc aacaaaacta gtcaaggtgt ccaagaagag catccaaggg tgttcctgca
     4021 aaggttggtg cgggaacaag caatgtggat gcagaaagca aaagtcagac tgtaatgtgt
     4081 cctgcagctg tgaccccaca aagtgtcgca accgccatca gaaccaggac aattcagatg
     4141 ctattgagct aaaccaagat tctgaaaact ctttcaaact ggaagatccc acagaggtga
     4201 cctcaggctt aagcttcttc caccctatct gtgcaactcc cagtagcaag atcctaaaag
     4261 aaatgtgtga tgcagatcag gtgcagctga agcagcccgt gtttgtctct tcttctgacc
     4321 acccagaact gaagtctata gcgtcagaat cacaagaaaa taaggccata gggaagaaga
     4381 agaagcgagc tctggccagc aataccagct tcttctctgg ctgctctcct atccaagaag
     4441 agtcccacgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     4501 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     4561 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4621 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4681 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4741 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4801 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4861 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4921 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4981 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5041 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5101 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     5161 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5221 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5281 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5341 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5401 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     5461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5521 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5581 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     5641 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5761 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     5821 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     5881 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5941 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     6001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     6061 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6121 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     6181 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     6241 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6301 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6361 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6421 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6481 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6541 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6601 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6661 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6721 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6781 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6841 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6901 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6961 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7021 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7081 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     7141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     7201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     7261 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7381 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7441 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7501 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     8041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     8281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8521 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9541 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     9601 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     9661 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     9721 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
