LOCUS       sequence-123466-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cerulean-tagged, kinase-dead Plk1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  TITLE     pCer-C3-Plk1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6501)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCer-C3-Plk1-K82R
FEATURES             Location/Qualifiers
     source          1..6501
                     /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 monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     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"
     polyA_signal    3289..3410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3326..3345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3380..3399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3417..3872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3554..3575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3766..3785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3899..4003
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4005..4362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4005..4196
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4028..4048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4213..4348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4397..5191
                     /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             4397..5191
                     /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(4451..4470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5061..5080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5379..5398)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5423..5470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5799..6387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6288..6307
                     /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 tggggcgtgc agtgcttcgc 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 gtacaacgcc atcagcgaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt actcagatct cgagctcaag cttcgaattc taagtgctgc agtgactgca
     1381 gggaagctgg cacgggcacc ggccgaccct gggaaagccg gggtccccgg agttgcagct
     1441 cccggagctc cggcggcggc tccaccggcg aaagagatcc cggaggtcct agtggaccca
     1501 cgcagccggc ggcgctatgt gcggggccgc tttttgggca agggcggctt tgccaagtgc
     1561 ttcgagatct cggacgcgga caccaaggag gtgttcgcgg gcaggattgt gcctaagtct
     1621 ctgctgctca agccgcacca gagggagaag atgtccatgg aaatatccat tcaccgcagc
     1681 ctcgcccacc agcacgtcgt aggattccac ggctttttcg aggacaacga cttcgtgttc
     1741 gtggtgttgg agctctgccg ccggaggtct ctcctggagc tgcacaagag gaggaaagcc
     1801 ctgactgagc ctgaggcccg atactaccta cggcaaattg tgcttggctg ccagtacctg
     1861 caccgaaacc gagttattca tcgagacctc aagctgggca accttttcct gaatgaagat
     1921 ctggaggtga aaatagggga ttttggactg gcaaccaaag tcgaatatga cggggagagg
     1981 aagaagaccc tgtgtgggac tcctaattac atagctcccg aggtgctgag caagaaaggg
     2041 cacagtttcg aggtggatgt gtggtccatt gggtgtatca tgtatacctt gttagtgggc
     2101 aaaccacctt ttgagacttc ttgcctaaaa gagacctacc tccggatcaa gaagaatgaa
     2161 tacagtattc ccaagcacat caaccccgtg gccgcctccc tcatccagaa gatgcttcag
     2221 acagatccca ctgcccgccc aaccattaac gagctgctta atgacgagtt ctttacttct
     2281 ggctatatcc ctgcccgtct ccccatcacc tgcctgacca ttccaccaag gttttcgatt
     2341 gctcccagca gcctggaccc cagcaaccgg aagcccctca cagtcctcaa taaaggcttg
     2401 gagaaccccc tgcctgagcg tccccgggaa aaagaagaac cagtggttcg agagacaggt
     2461 gaggtggtcg actgccacct cagtgacatg ctgcagcagc tgcacagtgt caatgcctcc
     2521 aagccctcgg agcgtgggct ggtcaggcaa gaggaggctg aggatcctgc ctgcatcccc
     2581 atcttctggg tcagcaagtg ggtggactat tcggacaagt acggccttgg gtatcagctc
     2641 tgtgataaca gcgtgggggt gctcttcaat gactcaacac gcctcatcct ctacaatgat
     2701 ggtgacagcc tgcagtacat agagcgtgac ggcactgagt cctacctcac cgtgagttcc
     2761 catcccaact ccttgatgaa gaagatcacc ctccttaaat atttccgcaa ttacatgagc
     2821 gagcacttgc tgaaggcagg tgccaacatc acgccgcgcg aaggtgatga gctcgcccgg
     2881 ctgccctacc tacggacctg gttccgcacc cgcagcgcca tcatcctgca cctcagcaac
     2941 ggcagcgtgc agatcaactt cttccaggat cacaccaagc tcatcttgtg cccactgatg
     3001 gcagccgtga cctacatcga cgagaagcgg gacttccgca cataccgcct gagtctcctg
     3061 gaggagtacg gctgctgcaa ggagctggcc agccggctcc gctacgcccg cactatggtg
     3121 gacaagctgc tgagctcacg ctcggccagc aaccgtctca aggcctccta agctagataa
     3181 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3241 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     3421 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3481 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3541 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3601 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3661 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3721 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3781 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3841 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3901 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3961 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4021 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4081 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4141 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4201 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4261 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4321 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4381 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4441 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4501 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4561 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4621 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4681 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4741 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4801 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4861 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4921 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4981 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5041 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5101 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5161 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5221 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5281 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5341 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5401 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5461 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5521 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5581 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5641 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5701 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5761 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5821 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5881 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5941 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6001 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6061 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6121 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6181 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6241 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6301 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6361 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6421 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6481 aaccgtatta ccgccatgca t
//
