LOCUS       sequence-202165-        6155 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 6155)
  AUTHORS   Gavet O, Pines J
  TITLE     Progressive activation of CyclinB1-Cdk1 coordinates entry to 
            mitosis.
  JOURNAL   Dev Cell. 2010 Apr 20. 18(4):533-43.
  PUBMED    20412769
REFERENCE   2  (bases 1 to 6155)
  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: "Dev Cell. 
            2010 Apr 20. 18(4):533-43."
COMMENT     Sequence Label: cdk1 FRET sensor (2327)
FEATURES             Location/Qualifiers
     source          1..6155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(600..602,603..605,606..1316)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     CDS             join(2082..2084,2085..2087,2088..2798)
                     /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     2735..2756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2799..2822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2955..3076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2992..3011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3046..3065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3083..3538)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3220..3241)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3432..3451
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3565..3669
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3694..3714)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3738..3948
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3760..3956
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3807..3942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3991..4785
                     /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             3991..4785
                     /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(4045..4064)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4655..4674
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4973..4992)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5017..5064
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5393..5981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5882..5901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgctagc gctaccggta
      601 tggtgagcaa aggcgaagag ctgttcaccg gcgtggtgcc catcctggtg gagctggacg
      661 gcgacgtgaa cggccacaag ttcagcgtga gcggcgaggg cgagggcgac gccacctacg
      721 gcaagctgac cctgaagctg ctgtgcacca ccggcaagct gcccgtgccc tggcccaccc
      781 tggtgaccac cctgggctac ggcgtgcagt gcttcgcccg gtaccccgac cacatgaagc
      841 agcacgactt cttcaagagc gccatgcccg agggctacgt gcaggagcgg accatcttct
      901 tcaaggacga cggcaactac aagacccggg ccgaggtgaa gttcgagggc gacaccctgg
      961 tgaaccggat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctgggccaca
     1021 agctggagta caactacaac agccacaacg tgtacatcac cgccgacaag cagaagaacg
     1081 gcatcaaggc caacttcaag atccggcaca acatcgagga cggcggcgtg cagctggccg
     1141 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1201 acctgagcta ccagagcgcc ctgttcaagg accccaacga gaagcgggac cacatggtgc
     1261 tgctggagtt cctgaccgcc gccggcatca ccgagggcat gaacgagctc tataagagat
     1321 ctcacctcag tgacatgctg cagcagctgc acagtgtcaa tgcctccaag ccctcggagc
     1381 gtgggctggt caggcaagag gaggctgagg atcctgcctg catccccatc ttctgggtca
     1441 gcaagtgggt ggactattcg gacaagtacg gccttgggta tcagctctgt gataacagcg
     1501 tgggggtgct cttcaatgac tcaacacgcc tcatcctcta caatgatggt gacagcctgc
     1561 agtacataga gcgtgacggc actgagtcct acctcaccgt gagttcccat cccaactcct
     1621 tgatgaagaa gatcaccctc cttaaatatt tccgcaatta catgagcgag cacttgctga
     1681 aggcaggtgc caacatcacg ccgcgcgaag gtgatgagct cgcccggctg ccctacctac
     1741 ggacctggtt ccgcacccgc agcgccatca tcctgcacct cagcaacggc agcgtgcaga
     1801 tcaacttctt ccaggatcac accaagctca tcttgtgccc actgatggca gccgtgacct
     1861 acatcgacga gaagcgggac ttccgcacat accgcctgag tctcctggag gagtacggct
     1921 gctgcaagga gctggccagc cggctccgct acgcccgcac tatggtggac aagctgctga
     1981 gcggcggcac aggcggcaca ggcggcacag gcggcacagg cggtacccct gagcctattt
     2041 tggttgatac ttcctctcca agcccaatgg aaacactcga gatggtgagc aagggcgagg
     2101 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2161 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2221 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2281 ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2341 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2401 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2461 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacgcca
     2521 tcagcgacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     2581 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2641 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     2701 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2761 ccgccgggat cactctcggc atggacgagc tgtacaagga ttacaaggat gacgacgata
     2821 agtaaggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2881 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2941 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3061 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3121 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3181 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3241 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3301 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3361 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3421 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3481 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3661 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3721 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3781 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3961 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4021 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4081 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4141 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4201 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4261 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4321 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4381 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4441 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4501 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4561 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4621 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4681 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4741 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4801 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4861 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4921 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4981 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5041 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5101 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5161 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5221 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5281 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5341 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5401 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5461 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5521 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5581 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5641 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5701 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5761 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5821 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5881 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5941 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6001 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6061 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     6121 gggtcattag ttcatagccc atatatggag ttccg
//
