LOCUS       V022839                 8365 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022839
VERSION     V022839
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             890..3799
                     /label="PLK4(NM_014264)"
                     /note="PLK4(NM_014264)"
                     /gene="PLK4"
     CDS             3800..3829
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     polyA_signal    3932..4156
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4202..4630
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4644..4973
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5040..5834
                     /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    6008..6129
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6178..6194)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6202..6218
                     /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(6226..6256)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6271..6292
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6580..7165)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7336..8196)
                     /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(8197..8301)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtacca tggcgacctg
      901 catcggggag aagatcgagg attttaaagt tggaaatctg cttggtaaag gatcatttgc
      961 tggtgtctac agagctgagt ccattcacac tggtttggaa gttgcaatca aaatgataga
     1021 taagaaagcc atgtacaaag caggaatggt acagagagtc caaaatgagg tgaaaataca
     1081 ttgccaattg aaacatcctt ctatcttgga gctttataac tattttgaag atagcaatta
     1141 tgtgtatctg gtattagaaa tgtgccataa tggagaaatg aacaggtatc taaagaatag
     1201 agtgaaaccc ttctcagaaa atgaagctcg acacttcatg caccagatca tcacagggat
     1261 gttgtatctt cattctcatg gtatactaca ccgggacctc acactttcta acctcctact
     1321 gactcgtaat atgaacatca agattgctga ttttgggctg gcaactcaac tgaaaatgcc
     1381 acatgaaaag cactatacat tatgtggaac tcctaactac atttcaccag aaattgccac
     1441 tcgaagtgca catggccttg aatctgatgt ttggtccctg ggctgtatgt tttatacatt
     1501 acttatcggg agaccaccct tcgacactga cacagtcaag aacacattaa ataaagtagt
     1561 attggcagat tatgaaatgc catctttttt gtcaatagag gccaaggacc ttattcacca
     1621 gttacttcgt agaaatccag cagatcgttt aagtctgtct tcagtattgg accatccttt
     1681 tatgtcccga aattcttcaa caaaaagtaa agatttagga actgtggaag actcaattga
     1741 tagtgggcat gccacaattt ctactgcaat tacagcttct tccagtacca gtataagtgg
     1801 tagtttattt gacaaaagaa gacttttgat tggtcagcca ctcccaaata aaatgactgt
     1861 atttccaaag aataaaagtt caactgattt ttcttcttca ggagatggaa acagttttta
     1921 tactcagtgg ggaaatcaag aaaccagtaa tagtggaagg ggaagagtaa ttcaagatgc
     1981 agaagaaagg ccacattctc gataccttcg tagagcttat tcctctgata gatctggcac
     2041 ttctaatagt cagtctcaag caaaaacata tacaatggaa cgatgtcact cagcagaaat
     2101 gctttcagtg tccaaaagat caggaggagg tgaaaatgaa gagaggtact cacccacaga
     2161 caacaatgcc aacattttta acttctttaa agaaaagaca tccagtagtt ctggatcttt
     2221 tgaaagacct gataacaatc aagcactctc caatcatctt tgtccaggaa aaactccttt
     2281 tccatttgca gacccgacac ctcagactga aaccgtacaa cagtggtttg ggaatctgca
     2341 aataaatgct catttaagaa aaactactga atatgacagc atcagcccaa accgggactt
     2401 ccagggccat ccagatttgc agaaggacac atcaaaaaat gcctggactg atacaaaagt
     2461 caaaaagaac tctgatgctt ctgataatgc acattctgta aaacagcaaa ataccatgaa
     2521 atatatgact gcacttcaca gtaaacctga gataatccaa caagaatgtg tttttggctc
     2581 agatcctctt tctgaacaga gcaagactag gggtatggag ccaccatggg gttatcagaa
     2641 tcgtacatta agaagcatta catctccgtt ggttgctcac aggttaaaac caatcagaca
     2701 gaaaaccaaa aaggctgtgg tgagcatact tgattcagag gaggtgtgtg tggagcttgt
     2761 aaaggagtat gcatctcaag aatatgtgaa agaagttctt cagatatcta gtgatggaaa
     2821 tacgatcact atttattatc caaatggtgg tagaggtttt cctcttgctg atagaccacc
     2881 ctcacctact gacaacatca gtaggtacag ctttgacaat ttaccagaaa aatactggcg
     2941 aaaatatcaa tatgcttcca ggtttgtaca gcttgtaaga tctaaatctc ccaaaatcac
     3001 ttattttaca agatatgcta aatgcatttt gatggagaat tctcctggtg ctgattttga
     3061 ggtttggttt tatgatgggg taaaaataca caaaacagaa gatttcattc aggtgattga
     3121 aaagacaggg aagtcttaca ctttaaaaag tgaaagtgaa gttaatagct tgaaagagga
     3181 gataaaaatg tatatggacc atgctaatga gggtcatcgt atttgtttag cactggaatc
     3241 cataatttca gaagaggaaa ggaaaactag gagtgctccc tttttcccaa taatcatagg
     3301 aagaaaacct ggtagtacta gttcacctaa ggccttatca cctcctcctt ctgtggattc
     3361 aaattaccca acgagagaga gagcatcttt caacagaatg gtcatgcata gtgctgcttc
     3421 tccaacacag gcaccaatcc ttaatccctc tatggttaca aatgaaggac ttggtcttac
     3481 aactacagct tctggaacag acatctcttc taatagtcta aaagattgtc ttcctaaatc
     3541 agcacaactt ttgaaatctg tttttgtgaa aaatgttggt tgggctacac agttaactag
     3601 tggagctgtg tgggttcagt ttaatgatgg gtcccagttg gttgtgcagg caggagtgtc
     3661 ttctatcagt tatacctcac caaatggtca aacaactagg tatggagaaa atgaaaaatt
     3721 accagactac atcaaacaga aattacagtg tctgtcttcc atccttttga tgttttctaa
     3781 tccgactcct aattttcatg agcagaaact catctctgaa gaggatctgt gaggatccac
     3841 tagtccagtg tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     3901 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3961 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4021 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4081 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4141 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4201 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4261 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4321 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4381 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4441 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4501 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4561 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4621 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4681 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4801 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4861 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4921 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4981 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5041 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5101 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5161 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5221 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5281 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5341 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5401 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5461 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5521 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5581 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5641 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5701 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5761 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5821 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5881 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5941 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6001 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6121 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6181 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6241 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6301 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6361 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6421 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6481 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6541 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6601 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6661 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6721 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6781 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6841 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6901 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6961 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7021 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7081 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     7141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     8341 gggagatctc ccgatcccct atggt
//
