LOCUS       sequence-128985-         889 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a Golgi located FRB domain in mammalian cells (the ECFP is
            not fluorescent).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 889)
  AUTHORS   van Unen J, Reinhard NR, Yin T, Wu YI, Postma M, Gadella TW, 
            Goedhart J
  TITLE     Plasma membrane restricted RhoGEF activity is sufficient for 
            RhoA-mediated actin polymerization.
  JOURNAL   Sci Rep. 2015 Oct 5;5:14693. doi: 10.1038/srep14693.
  PUBMED    26435194
REFERENCE   2  (bases 1 to 889)
  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: "Sci Rep."; 
            date: "2015-10-5"; volume: "5:14693. doi"; pages: " 
            10.1038/srep14693"
COMMENT     Sequence Label: FRB-ECFP(W66A)-Giantin
FEATURES             Location/Qualifiers
     source          1..889
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      29..38
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             47..325
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             332..358
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      422..431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(428..430,431..433,434..889)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTAGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYIT"
     primer_bind     complement(473..494)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(734..753)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ttagtgaacc gtcagatccg ctagcgccgc caccatggct tctagaatcc tctggcatga
       61 gatgtggcat gaaggcctgg aagaggcatc tcgtttgtac tttggggaaa ggaacgtgaa
      121 aggcatgttt gaggtgctgg agcccttgca tgctatgatg gaacggggcc cccagactct
      181 gaaggaaaca tcctttaatc aggcctatgg tcgagattta atggaggccc aagagtggtg
      241 caggaagtac atgaaatcag ggaatgtcaa ggacctcctc caagcctggg acctctatta
      301 tcatgtgttc cgacgaatct caaagactag ttatccgtac gacgtaccag actacgcaag
      361 tgctggtagt gctggtagtg ctggtagtgc tggtagtgct ggtagtgctg gttcaccggt
      421 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      481 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      541 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      601 gcccaccctc gtgaccaccc tgaccgccgg cgtgcagtgc ttcagccgct accccgacca
      661 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      721 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      781 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      841 ggggcacaag ctggagtaca actacatcag ccacaacgtc tatatcacc
//
