LOCUS       sequence-265937-        7052 bp    DNA     circular SYN 15-DEC-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 7052)
  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 7052)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: EGFP-plk1-AKAP
FEATURES             Location/Qualifiers
     source          1..7052
                     /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    3840..3961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3877..3896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3931..3950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3968..4423)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4105..4126)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4317..4336
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4450..4554
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4556..4913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4556..4747
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4579..4599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4764..4899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4948..5742
                     /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             4948..5742
                     /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(5002..5021)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5612..5631
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5930..5949)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5974..6021
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /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 gaagaccctg 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 caacattgaa gccatcattg cctctgaaaa agaagtatgg
     3241 aacagagaaa aattgactct ccagaaatct ttgaaaaggg cagaggctga agtatacaaa
     3301 ctgaaagctg aactaagaaa tgactcttta cttcaaactc tgagccctga ttctgaacat
     3361 gtcactttaa agagaattta tggtaaatac ttgagggcag aaagttttcg aaaggctctc
     3421 atttaccaga agaaatacct gctgctgtta ctgggtgggt tccaggaatg tgaagatgcc
     3481 accttggccc tgcttgcccg gatggggggg cagccagctt tcacggatct agaggtgatc
     3541 accaatcgcc caaagggctt caccaggttt cggtcggccg tcagagtatc cattgcaatt
     3601 tccagaatga aatttttggt tcgacggtgg catcgagtca caggttctgt ttccatcaat
     3661 attaacagag atggctttgg actgaatcaa ggtgcataac cgcgggcccg ggatccaccg
     3721 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3781 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3961 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4021 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4081 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4141 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4201 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4261 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4321 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4381 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4441 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4501 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4561 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4621 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4681 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4741 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4801 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4861 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4921 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4981 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5041 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5101 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     5161 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5221 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5281 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5341 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5401 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5461 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5521 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5581 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5641 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5701 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5761 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5821 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5881 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5941 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     6001 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     6061 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     6121 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     6181 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgccatgc at
//
