LOCUS       sequence-247210-        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry PLK1 in mammalian cells and zebrafish embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  AUTHORS   Colicino EG, Stevens K, Curtis E, Rathbun L, Bates M, Manikas J, 
            Amack J, Freshour J, Hehnly H
  TITLE     Chromosome misalignment is associated with PLK1 activity at 
            cenexin-positive mitotic centrosomes.
  JOURNAL   Mol Biol Cell. 2019 Jun 15;30(13):1598-1609. doi: 
            10.1091/mbc.E18-12-0817. Epub 2019 May 1.
  PUBMED    31042116
REFERENCE   2  (bases 1 to 6619)
  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: 
            "10.1091/mbc.E18-12-0817"; journalName: "Mol Biol Cell"; date: 
            "2019-06-15- 15"; volume: "30"; issue: "13"; pages: "1598-1609"
COMMENT     Sequence Label: pCS2-PLK1-mCherry
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1908..2618
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2053..2071)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2321..2340
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2671..2805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2695..2714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2749..2768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2918..2938)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2918..2936)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2957..2973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2957..2973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2970..2992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2981..2997
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3005..3011,3012..3029,3030..3035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3050..3071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3188..3205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3359..3947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3439..3458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4118..4909,4910..4978))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4741..4760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4979..5083)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5109..5564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5246..5267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5458..5477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5635..6619
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagt ctcgagacca tgagtgctgc agtgactgca gggaagctgg
      121 cacgggcacc ggccgaccct gggaaagccg gggtccccgg agttgcagct cccggagctc
      181 cggcggcggc tccaccggcg aaagagatcc cggaggtcct agtggaccca cgcagccggc
      241 ggcgctatgt gcggggccgc tttttgggca agggcggctt tgccaagtgc ttcgagatct
      301 cggacgcgga caccaaggag gtgttcgcgg gcaagattgt gcctaagtct ctgctgctca
      361 agccgcacca gagggagaag atgtccatgg aaatatccat tcaccgcagc ctcgcccacc
      421 agcacgtcgt aggattccac ggctttttcg aggacaacga cttcgtgttc gtggtgttgg
      481 agctctgccg ccggaggtct ctcctggagc tgcacaagag gaggaaagcc ctgactgagc
      541 ctgaggcccg atactaccta cggcaaattg tgcttggctg ccagtacctg caccgaaacc
      601 gagttattca tcgagacctc aagctgggca accttttcct gaatgaagat ctggaggtga
      661 aaatagggga ttttggactg gcaaccaaag tcgaatatga cggggagagg aagaagaccc
      721 tgtgtgggac tcctaattac atagctcccg aggtgctgag caagaaaggg cacagtttcg
      781 aggtggatgt gtggtccatt gggtgtatca tgtatacctt gttagtgggc aaaccacctt
      841 ttgagacttc ttgcctaaaa gagacctacc tccggatcaa gaagaatgaa tacagtattc
      901 ccaagcacat caaccccgtg gccgcctccc tcatccagaa gatgcttcag acagatccca
      961 ctgcccgccc aaccattaac gagctgctta atgacgagtt ctttacttct ggctatatcc
     1021 ctgcccgtct ccccatcacc tgcctgacca ttccaccaag gttttcgatt gctcccagca
     1081 gcctggaccc cagcaaccgg aagcccctca cagtcctcaa taaaggcttg gagaaccccc
     1141 tgcctgagcg tccccgggaa aaagaagaac cagtggttcg agagacgggt gaggtggtcg
     1201 actgccacct cagtgacatg ctgcagcagc tgcacagtgt caatgcctcc aagccctcgg
     1261 agcgtgggct ggtcaggcaa gaggaggctg aggatcctgc ctgcatcccc atcttctggg
     1321 tcagcaagtg ggtggactat tcggacaagt acggccttgg gtatcagctc tgtgataaca
     1381 gcgtgggggt gctcttcaat gactcaacac gcctcatcct ctacaatgat ggtgacagcc
     1441 tgcagtacat agagcgtgac ggcactgagt cctacctcac cgtgagttcc catcccaact
     1501 ccttgatgaa gaagatcacc ctccttaaat atttccgcaa ttacatgagc gagcacttgc
     1561 tgaaggcagg tgccaacatc acgccgcgcg aaggtgatga gctcgcccgg ctgccctacc
     1621 tacggacctg gttccgcacc cgcagcgcca tcatcctgca cctcagcaac ggcagcgtgc
     1681 agatcaactt cttccaggat cacaccaagc tcatcttgtg cccactgatg gcagccgtga
     1741 cctacatcga cgagaagcgg gacttccgca cataccgcct gagtctcctg gaggagtacg
     1801 gctgctgcaa ggagctggcc agccggctcc gctacgcccg cactatggtg gacaagctgc
     1861 tgagctcacg ctcggccagc aaccgtctca aggcctcccg aattctgatg gtgagcaagg
     1921 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     1981 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2041 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2101 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2161 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2221 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2281 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2341 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2401 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2461 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2521 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2581 gccactccac cggcggcatg gacgagctgt acaagtaaga attcgccagc acagtctgca
     2641 gtctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     2701 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2761 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2821 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
     2881 gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
     2941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3001 cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3781 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4081 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4201 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4261 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4321 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4561 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4621 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4801 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4981 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5041 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5101 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5161 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     5221 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     5281 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5341 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5401 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5461 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5521 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     5581 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
     5641 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
     5701 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
     5761 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
     5821 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
     5881 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
     5941 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
     6001 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
     6061 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
     6121 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
     6181 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
     6241 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
     6301 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
     6361 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
     6421 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
     6481 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
     6541 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     6601 caatgctcgt ttagggaac
//
