LOCUS       sequence-265935-        6545 bp    DNA     circular SYN 14-APR-2025
DEFINITION  To study the processes involved in early mitotic events, mitotic 
            progression, and control of re-entry after DNA damage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6545)
  AUTHORS   Kishi K, van Vugt MA, Okamoto K, Hayashi Y, Yaffe MB
  TITLE     Functional dynamics of Polo-like kinase 1 at the centrosome.
  JOURNAL   Mol Cell Biol. 2009 Jun;29(11):3134-50. doi: 10.1128/MCB.01663-08. 
            Epub 2009 Mar 23.
  PUBMED    19307309
REFERENCE   2  (bases 1 to 6545)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/MCB.01663-08"; journalName: "Mol Cell Biol"; date: 
            "2009-06"; volume: "29"; issue: "11"; pages: "3134-50"
COMMENT     Sequence Label: plk1 T210A
FEATURES             Location/Qualifiers
     source          1..6545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1351..1380
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3333..3454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3370..3389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3424..3443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3461..3916)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3598..3619)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3810..3829
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3943..4047
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4049..4406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4049..4240
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4072..4092)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4257..4392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4441..5235
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4441..5235
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4495..4514)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5105..5124
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5423..5442)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5467..5514
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5843..6431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6332..6351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgaggt gaacaaaaac tcatctcaga agaggatctg
     1381 agtgctgcag tgactgcagg gaagctggca cgggcaccgg ccgaccctgg gaaagccggg
     1441 gtccccggag ttgcagctcc cggagctccg gcggcggctc caccggcgaa agagatcccg
     1501 gaggtcctag tggacccacg cagccggcgg cgctatgtgc ggggccgctt tttgggcaag
     1561 ggcggctttg ccaagtgctt cgagatctcg gacgcggaca ccaaggaggt gttcgcgggc
     1621 aagattgtgc ctaagtctct gctgctcaag ccgcaccaga gggagaagat gtccatggaa
     1681 atatccattc accgcagcct cgcccaccag cacgtcgtag gattccacgg ctttttcgag
     1741 gacaacgact tcgtgttcgt ggtgttggag ctctgccgcc ggaggtctct cctggagctg
     1801 cacaagagga ggaaagccct gactgagcct gaggcccgat actacctacg gcaaattgtg
     1861 cttggctgcc agtacctgca ccgaaaccga gttattcatc gagacctcaa gctgggcaac
     1921 cttttcctga atgaagatct ggaggtgaaa ataggggatt ttggactggc aaccaaagtc
     1981 gaatatgacg gggagaggaa gaaggccctg tgtgggactc ctaattacat agctcccgag
     2041 gtgctgagca agaaagggca cagtttcgag gtggatgtgt ggtccattgg gtgtatcatg
     2101 tataccttgt tagtgggcaa accacctttt gagacttctt gcctaaaaga gacctacctc
     2161 cggatcaaga agaatgaata cagtattccc aagcacatca accccgtggc cgcctccctc
     2221 atccagaaga tgcttcagac agatcccact gcccgcccaa ccattaacga gctgcttaat
     2281 gacgagttct ttacttctgg ctatatccct gcccgtctcc ccatcacctg cctgaccatt
     2341 ccaccaaggt tttcgattgc tcccagcagc ctggacccca gcaaccggaa gcccctcaca
     2401 gtcctcaata aaggcttgga gaaccccctg cctgagcgtc cccgggaaaa agaagaacca
     2461 gtggttcgag agacaggtga ggtggtcgac tgccacctca gtgacatgct gcagcagctg
     2521 cacagtgtca atgcctccaa gccctcggag cgtgggctgg tcaggcaaga ggaggctgag
     2581 gatcctgcct gcatccccat cttctgggtc agcaagtggg tggactattc ggacaagtac
     2641 ggccttgggt atcagctctg tgataacagc gtgggggtgc tcttcaatga ctcaacacgc
     2701 ctcatcctct acaatgatgg tgacagcctg cagtacatag agcgtgacgg cactgagtcc
     2761 tacctcaccg tgagttccca tcccaactcc ttgatgaaga agatcaccct ccttaaatat
     2821 ttccgcaatt acatgagcga gcacttgctg aaggcaggtg ccaacatcac gccgcgcgaa
     2881 ggtgatgagc tcgcccggct gccctaccta cggacctggt tccgcacccg cagcgccatc
     2941 atcctgcacc tcagcaacgg cagcgtgcag atcaacttct tccaggatca caccaagctc
     3001 atcttgtgcc cactgatggc agccgtgacc tacatcgacg agaagcggga cttccgcaca
     3061 taccgcctga gtctcctgga ggagtacggc tgctgcaagg agctggccag ccggctccgc
     3121 tacgcccgca ctatggtgga caagctgctg agctcacgct cggccagcaa ccgtctcaag
     3181 gcctccaagg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     3241 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3301 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3421 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3481 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3541 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3601 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3661 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3721 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3781 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3841 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3901 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3961 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4021 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4321 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4381 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4441 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4501 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4561 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4621 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4681 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4741 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4801 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4861 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4921 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4981 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5041 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5101 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5161 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5221 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5281 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5341 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5401 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5461 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5521 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5581 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5641 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6541 tgcat
//
