LOCUS       V028519                 8490 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028519
VERSION     V028519
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1029..4637
                     /label="Nek1(NM_175089)"
                     /note="Nek1(NM_175089)"
                     /gene="Nek1"
     CDS             4656..4685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4704..4727
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4747..4763)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4774..5396
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6462..6509
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6741..7535)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(7570..7927)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(7929..8033)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8060..8488
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggagaagtat gtgagactgc agaagattgg agaaggttca tttggaaaag
     1081 ctgttcttgt taaatcgaca gaggatggca gacattatgt catcaaggaa attaacatct
     1141 caagaatgtc tgataaagaa aggcaagaat caaggagaga agttgctgta ttggcaaaca
     1201 tgaagcatcc aaatattgtc caatataaag aatcatttga agaaaatggc tctctctaca
     1261 tagtaatgga ttactgtgaa ggaggtgatt tgtttaaacg aataaatgct cagaaaggcg
     1321 ctctgtttca agaagaccag attttggact ggtttgtgca gatatgtttg gctctgaagc
     1381 atgtacatga tagaaaaatt cttcaccgag acataaagtc acagaacata tttctaacca
     1441 aagatgggac agtgcagctt ggagattttg gaattgctcg agttcttaat agtactgtag
     1501 agctggctcg aacttgcata ggcactccat actacttgtc acctgaaatc tgtgaaaaca
     1561 agccttataa caataaaagt gacatttggg ctttgggctg tgtcctttat gagttgtgta
     1621 cacttaaaca tgcatttgaa gctggaaaca tgaaaaacct ggtactgaag ataatctccg
     1681 gatcctttcc tccagtgtct ccacattact cctatgatct ccgcagcttg ctgtctcagt
     1741 tatttaaaag aaatcctagg gatagaccat cagtcaactc catattggag aaaggtttta
     1801 tagctaaacg aatcgaaaag tttctctccc ctcagcttat tgcagaagaa ttttgtctaa
     1861 aaacactttc aaagtttgga ccacagcctc tcccaggtaa aagaccagca tcaggacaag
     1921 gtgtcagttc ttttgtccct gctcagaaaa tcacaaagcc tgctgctaaa tacggagtgc
     1981 ctttaacata taagaagtat ggagataaaa agttacttga gaaaaaacca cccccaaaac
     2041 ataaacaggc ccatcaaatt cccgtgaaga aaatgaattc tggagaagaa aggaagaaaa
     2101 tgtctgagga agcagcaaaa aaaagaaggt tggaatttat tgagaaagaa aagaagcaaa
     2161 aggatcagat taggttcctg aaggctgagc agatgaagcg gcaagagaag cagcggttgg
     2221 agaggataaa tagggccagg gaacaaggat ggaggaatgt tttaagggct ggtggaagcg
     2281 gtgaagtaaa ggcttccttt tttggcattg gaggggctgt ctctccatca ccgtgttctc
     2341 ctcgaggcca gtatgaacat taccatgcca tttttgacca aatgcagcgg ctaagagcag
     2401 aagataatga agcaagatgg aaggggggaa tctatggtcg atggctccca gaaaggcaaa
     2461 aaggacactt agctgtagag agagccaacc aagtggaaga attcctacag cgtaaacgag
     2521 aagctatgca gaataaagcc cgagccgaag gacacgtggt ttatttggca agactgaggc
     2581 aaataagact acaaaatttt aatgagcgcc aacagattaa agccaaactt cgtggtgaga
     2641 ataaagaagc tgatggtacc aaaggacaag aagcaactga agagactgac atgaggctca
     2701 aaaagatgga gtcacttaag gcgcaaacaa atgcacgtgc tgctgtacta aaagaacagc
     2761 tggagcgaaa aagaaaggaa gcttatgaaa gagaaaagaa agtatgggaa gaacatttgg
     2821 tggcgagggt aaaaagctca gatgttcctc tgcctttgga acttcttgaa acaggtggtt
     2881 ctccatcaaa gcagcaggtg aagcctgtca tttctgtgac ttcagctttg aaagaagtgg
     2941 gcctggatgg aagtttaact gatacccagg aagaagaaat ggaaaagagt aacagtgcta
     3001 tttcaagtaa gcgagaaatc ctgcgtaggc taaatgaaaa tcttaaagct caagaggatg
     3061 aaaaggaaaa gcagcatcac tcaggttctt gtgagaccgt tggtcacaaa gatgagagag
     3121 agtatgagac agaaaatgcc atttcctctg atcgcaagaa gtgggagatg ggaggtcagc
     3181 ttgtgattcc tctcgatgca gtgacactgg atacatcctt ctctgcaacc gaaaaacata
     3241 ctgtgggaga ggttattaaa ttagattcta atggctctcc aagaaaagtc tgggggaaaa
     3301 accctacaga ttctgtgctg aagatacttg gagaagctga attacagcta cagacagaac
     3361 tactagaaaa cacatctttt aaaagtgagg tttatgctga agaggagaac tacaaaccct
     3421 tacttactga agaagagaat ctgcagtgca tttcaaaaga aataaatcca tcagctactg
     3481 ttgattctac tgaaacgaaa agtccaaagt ttactgaggt gtctccacaa atgtcagaag
     3541 gaaatgtgga agaacctgat gatttggaaa cagaagttct acaagagcca agtagcacac
     3601 acacagatgg gagtttgcca cctgttctta atgatgtgtg gactagagag aaggaagcag
     3661 ctaaggaaac tgagttggaa gataaggttg ctgtgcagca gagtgaagtt tgtgaagata
     3721 gaattccagg gaacgtggac caatcctgta aggatcagag agatcctgca gtagacgatt
     3781 ctccgcagtc tggctgtgat gtagagaagt cagtacagcc agaatcgatt ttccagaaag
     3841 tggttcattc taaggacttg aacttagttc aggcagttca ttgctcacca gaagaaccaa
     3901 ttccaattcg atctcactct gattctccac caaaaactaa gagcaagaat tccttactga
     3961 ttggactttc aactggtctg tttgatgcaa acaatccaaa gatgctgagg acctgctcac
     4021 ttccagatct ttccaagctg ttcagaaccc taatggacgt tcccactgtg ggggacgttc
     4081 atcaagacag tcttgaaatc gatgagctgg aagatgaacc aattaaagaa gggccttctg
     4141 attccgaaga cactgtattt gaagaaactg acacagattt acaagagctt caggcctcaa
     4201 tggagcagct gcttagggag caaccaggtg acgaatacag tgaggaggaa gagtctgttt
     4261 taaaaagcag cgatgtggag cagacagcaa gagggacaga tgccccagac gaggaggaca
     4321 accccagcag cgaaagcgcc ctgaacgagg aatggcactc agataatagt gacgctgaga
     4381 ccactagtga atgtgaatat gacagtgtct ttaaccattt agaggaacta agacttcact
     4441 tggagcaaga aatgggcttt gaaaagttct ttgaggttta tgagaaagta aaggctattc
     4501 atgaggatga agatgaaaat attgaaattt gttcaacaat agttgagaat attttgggca
     4561 atgagcacca gcatctctat gccaagattc tgcatttagt catggcagat ggagcctatc
     4621 aggaagataa tgatgaaacg cgtacgcggc cgctcgagca gaaactcatc tcagaagagg
     4681 atctggcagc aaatgatatc ctggattaca aggatgacga cgataaggtt taaacggccg
     4741 gccgcggtca tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca
     4801 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4861 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4921 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4981 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     5041 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     5101 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     5161 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     5221 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     5281 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     5341 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     5401 acgcggccag cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     6301 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     6361 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     6421 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     6481 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     6541 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     6601 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     6661 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     6721 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     6781 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     6841 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc
     6901 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     6961 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     7021 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     7081 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     7141 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     7201 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     7261 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     7321 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     7381 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     7441 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     7501 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     7561 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7621 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     7681 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     7741 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     7801 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     7861 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     7921 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8041 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     8161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8461 ctgatttaac aaaaatttaa cgcgaatttt
//
