LOCUS       sequence-80463-e        2024 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Please note this plasmid is discontinued. If you need this plasmid 
            for experiments, please directly contact Robert Campbell at 
            University of Alberta..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2024)
  AUTHORS   Belal AS, Sell BR, Hoi H, Davidson MW, Campbell RE
  TITLE     Optimization of a genetically encoded biosensor for cyclin B1-cyclin
            dependent kinase 1.
  JOURNAL   Mol Biosyst. 2014 Feb;10(2):191-5. doi: 10.1039/c3mb70402e.
  PUBMED    24281384
REFERENCE   2  (bases 1 to 2024)
  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"; journalName: 
            "Mol Biosyst"; date: "2014-02"; volume: "10"; issue: "2"; pages: 
            "191-5"
COMMENT     Sequence Label: cyclin B1-CDK1 FRET biosensor
FEATURES             Location/Qualifiers
     source          1..2024
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      4..13
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             22..729
                     /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(1288..1290,1291..1293,1294..2007)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1333..1354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1594..1613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1941..1962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gccgccacca tggcctcgag catggtgagc aagggcgagg agaccacaat gggcgtaatc
       61 aagcccgaca tgaagatcaa gctgaagatg gagggcaacg tgaatggcca cgccttcgtg
      121 atcgagggcg agggcgaggg caagccctac gacggcacca acaccatcaa cctggaggtg
      181 aaggagggag cccccctgcc cttctcctac gacattctga ccaccgcgtt cgcctacggc
      241 aacagggcct tcaccaagta ccccgacgac atccccaact acttcaagca gtccttcccc
      301 gagggctact cttgggagcg caccatgacc ttcgaggaca agggcatcgt gaaggtgaag
      361 tccgacatct ccatggagga ggactccttc atctacgaga tacacctcaa gggcgagaac
      421 ttccccccca acggccccgt gatgcagaag aagaccaccg gctgggacgc ctccaccgag
      481 aggatgtacg tgcgcgacgg cgtgctgaag ggcgacgtca agcacaagct gctgctggag
      541 ggcggcggcc accaccgcgt tgacttcaag accatctaca gggccaagaa ggcggtgaag
      601 ctgcccgact atcactttgt ggaccaccgc atcgagatcc tgaaccacga caaggactac
      661 aacaaggtga ccgtttacga gagcgccgtg gcccgcaact ccaccgacgg catggacgag
      721 ctgtacaagc ctaggttaac tttaaaacca ttgcctgaca gcattattca agaaagcctg
      781 gagattcagc aaggtgtgaa tccatttttc attggtagat ccgaggattg caattgtaaa
      841 attgaagaca ataggttgtc tcgagttcat tgcttcattt tcaaaaagag gcatgctgta
      901 ggcaaaagca tgtatgaatc tccggcacaa ggtttagatg atatttggta ttgccacacc
      961 ggaactaacg tgagctattt aaataataac cgcatgatac agggtacgaa attcctttta
     1021 caagacggag atgaaatcaa gatcatttgg gataaaaaca ataaatttgt cattggcttt
     1081 aaagtggaaa ttaacgatac tacaggtctg tttaacgagg gattaggtat gttacaagaa
     1141 caaagagtag tacttaagca aacagccgaa gaaaaagatt tggtgaaaaa gttagtcgac
     1201 ggcggcagcg gcggcggcgg cagcggcggc acccctgagc ctattttggt tgatacttcc
     1261 tctccaagcc caatggaaac aggtaccatg gtgagcaagg gcgaggagct gttcaccggg
     1321 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1381 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1441 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     1501 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1561 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1621 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1681 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1741 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1801 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1861 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     1921 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1981 ctcggcatgg acgagctgta caagtaaaag ctttaacctg cagg
//
