LOCUS       sequence-62959-e        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Liu D, Davydenko O, Lampson MA
  TITLE     Polo-like kinase-1 regulates kinetochore-microtubule dynamics and 
            spindle checkpoint silencing.
  JOURNAL   J Cell Biol. 2012 Aug 20;198(4):491-9. doi: 10.1083/jcb.201205090.
  PUBMED    22908307
REFERENCE   2  (bases 1 to 7871)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2012-08-20- 20"; volume: "198"; 
            issue: "4"; pages: "491-9"
COMMENT     Sequence Label: N-term Hec1 Plk1 FRET sensor c-jun substrate
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..90
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      155..164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             161..868
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(1412..1414,1415..2125)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     primer_bind     complement(1454..1475)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1715..1734)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4210..4331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4247..4266)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4301..4320
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4338..4793)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4475..4496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4687..4706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4820..4924
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4926..5283
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4926..5117
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4949..4969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5134..5269
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5318..6112
                     /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             5318..6112
                     /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(5372..5391)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5982..6001
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6300..6319)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6344..6391
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6720..7308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7209..7228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7483..7786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7787..7871,1..119)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
       61 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      121 gtttagtgaa ccgtcagatc cgctagcctc gagcgccacc atggtgagca agggcgagga
      181 gaccacaatg ggcgtaatca agcccgacat gaagatcaag ctgaagatgg agggcaacgt
      241 gaatggccac gccttcgtga tcgagggcga gggcgagggc aagccctacg acggcaccaa
      301 caccatcaac ctggaggtga aggagggagc ccccctgccc ttctcctacg acattctgac
      361 caccgcgttc gcctacggca acagggcctt caccaagtac cccgacgaca tccccaacta
      421 cttcaagcag tccttccccg agggctactc ttgggagcgc accatgacct tcgaggacaa
      481 gggcatcgtg aaggtgaagt ccgacatctc catggaggag gactccttca tctacgagat
      541 acacctcaag ggcgagaact tcccccccaa cggccccgtg atgcagaaga agaccaccgg
      601 ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa
      661 gcacaagctg ctgctggagg gcggcggcca ccaccgcgtt gacttcaaga ccatctacag
      721 ggccaagaag gcggtgaagc tgcccgacta tcactttgtg gaccaccgca tcgagatcct
      781 gaaccacgac aaggactaca acaaggtgac cgtttacgag agcgccgtgg cccgcaactc
      841 caccgacggc atggacgagc tgtacaaggc tggtaccaag ggtaatggta ggtttttaac
      901 tttaaaacca ttgcctgaca gcattattca agaaagcctg gagattcagc aaggtgtgaa
      961 tccatttttc attggtagat ccgaggattg caattgtaaa attgaagaca gtaggttgtc
     1021 tcgagttcat tgcttcattt tcaaaaagag gcatgctgta ggcaaaagca tgtatgaatc
     1081 tccggcacaa ggtttagatg atatttggta ttgccacacc ggaactaacg tgagctattt
     1141 aaataataac cgcatgatac agggtacgaa attcctttta caagacggag atgaaatcaa
     1201 gatcatttgg gataaaaaca ataaatttgt cattggcttt aaagtggaaa ttaacgatac
     1261 tacaggtctg tttaacgagg gattaggtat gttacaagaa caaagagtag tacttaagca
     1321 aacagccgaa gaaaaagatt tggtgaaaaa gttaggagga tccgacgacg ccctgaacgc
     1381 caccttcctg ccctccgagg gctccggagg agtgagcaag ggcgaggagc tgttcaccgg
     1441 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1501 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctgc tctgcaccac
     1561 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg
     1621 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1681 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1741 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1801 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1861 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1921 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1981 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgttcaaaga
     2041 ccccaacgag aagcgcgatc acatggtcct gctggagttc ctgaccgccg ccgggatcac
     2101 tgaaggcatg aacgagctgt acaagcgagc tcaagcttcg aattcaggcc ggcccatgaa
     2161 gcgcagttca gtttccagcg gtggtgctgg ccgcctctcc atgcaggagt taagatccca
     2221 ggatgtaaat aaacaaggcc tctatacccc tcaaaccaaa gagaaaccaa cctttggaaa
     2281 gttgagtata aacaaaccga catctgaaag aaaagtctcg ctatttggca aaagaactag
     2341 tggacatgga tcccggaata gtcaacttgg tatattttcc agttctgaga aaatcaagga
     2401 cccgagacca cttaatgaca aagcattcat tcagcagtgt attcgacaac tctgtgagtt
     2461 tcttacagaa aatggttatg cacataatgt gtccatgaaa tctctacaag ctccctctgt
     2521 taaagacttc ctgaagatct tcacatttct ttatggcttc ctgtgcccct catacgaact
     2581 tcctgacaca aagtttgaag aagaggttcc aagaatcttt aaagaccttg ggtatccttt
     2641 tgcactatcc aaaagctcca tgtacacagt gggggctcct catacatggc ctcacattgt
     2701 ggcagcctta gtttggctaa tagactgcat caagatacat actgccatga aagaaagctc
     2761 acctttattt gatgatgggc agccttgggg agaagaaact gaagatggaa ttatgcataa
     2821 taagttgttt ttggactaca ccataaaatg ctatgagagt tttatgagtg gtgccgacag
     2881 ctttgatgag atgaatgcag agctgcagtc aaaactgaag gatttattta atgtggatgc
     2941 ttttaagctg gaatcattag aagcaaaaaa cagagcattg aatgaacaga ttgcaagatt
     3001 ggaacaagaa agagaaaaag aaccgaatcg tctagagtcg ttgagaaaac tgaaggcttc
     3061 cttacaagga gatgttcaaa agtatcaggc atacatgagc aatttggagt ctcattcagc
     3121 cattcttgac cagaaattaa atggtctcaa tgaggaaatt gctagagtag aactagaatg
     3181 tgaaacaata aaacaggaga acactcgact acagaatatc attgacaacc agaagtactc
     3241 agttgcagac attgagcgaa taaatcatga aagaaatgaa ttgcagcaga ctattaataa
     3301 attaaccaag gacctggaag ctgaacaaca gaagttgtgg aatgaggagt taaaatatgc
     3361 cagaggcaaa gaagcgattg aaacacaatt agcagagtat cacaaattgg ctagaaaatt
     3421 aaaacttatt cctaaaggtg ctgagaattc caaaggttat gactttgaaa ttaagtttaa
     3481 tcccgaggct ggtgccaact gccttgtcaa atacagggct caagtttatg tacctcttaa
     3541 ggaactcctg aatgaaactg aagaagaaat taataaagcc ctaaataaaa aaatgggttt
     3601 ggaggatact ttagaacaat tgaatgcaat gataacagaa agcaagagaa gtgtgagaac
     3661 tctgaaagaa gaagttcaaa agctggatga tctttaccaa caaaaaatta aggaagcaga
     3721 ggaagaggat gaaaaatgtg ccagtgagct tgagtccttg gagaaacaca agcacctgct
     3781 agaaagtact gttaaccagg ggctcagtga agctatgaat gaattagatg ctgttcagcg
     3841 ggaataccaa ctagttgtgc aaaccacgac tgaagaaaga cgaaaagtgg gaaataactt
     3901 gcaacgtctg ttagagatgg ttgctacaca tgttgggtct gtagagaaac atcttgagga
     3961 gcagattgct aaagttgata gagaatatga agaatgcatg tcagaagatc tctcggaaaa
     4021 tattaaagag attagagata agtatgagaa gaaagctact ctaattaagt cttctgaaga
     4081 atgaggcgcg cctctagata actgatcata atcagccata ccacatttgt agaggtttta
     4141 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4201 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4261 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4321 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4381 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4441 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4501 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4561 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4621 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4681 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4741 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4801 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4861 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4921 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4981 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5041 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5101 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5161 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5221 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5281 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5341 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5401 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5461 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     5521 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5581 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5641 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5701 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5761 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5821 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5881 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5941 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6001 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6061 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6121 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6181 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6241 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6301 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     6361 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     6421 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     6481 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     6541 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     6601 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6661 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6721 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6781 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6841 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     6901 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6961 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7021 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7081 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7141 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7201 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7261 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7321 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7381 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
     7441 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     7501 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7561 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7621 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     7681 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     7741 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     7801 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     7861 cattgacgtc a
//
