LOCUS       sequence-149632-         722 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  FRET biosensor. To monitor Src kinase activity in mammalian cells by
            FRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 722)
  AUTHORS   Ouyang M, Sun J, Chien S, Wang Y
  TITLE     Determination of hierarchical relationship of Src and Rac at 
            subcellular locations with FRET biosensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Sep 23;105(38):14353-8. doi: 
            10.1073/pnas.0807537105. Epub 2008 Sep 17.
  PUBMED    18799748
REFERENCE   2  (bases 1 to 722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.0807537105"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2008-09-23- 23"; volume: "105"; issue: "38"; pages: "14353-8"
COMMENT     Sequence Label: Kras-Src FRET biosensor
FEATURES             Location/Qualifiers
     source          1..722
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     28..47
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        28..46
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      66..75
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             87..104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             108..140
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(144..152,153..167)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(174..176,177..179,180..692)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     primer_bind     complement(219..240)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(480..499)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
       61 cggccgccac catggtgcgg ggttctcatc atcatcatca tcatggtatg gctagcatga
      121 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat cccatggtga
      181 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      241 taaacggcca caggttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      301 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      361 ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      421 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgtaccatc ttcttcaagg
      481 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      541 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      601 agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca
      661 aggcccactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
      721 ac
//
