LOCUS       sequence-128655-         894 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a membrane 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 894)
  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 894)
  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: Lck-FRB-ECFP(W66A)
FEATURES             Location/Qualifiers
     source          1..894
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(74..91)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             106..384
                     /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             391..417
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(433..435,436..894)
                     /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="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTAGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITA"
     primer_bind     complement(475..496)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(736..755)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tgaaccgtca gatccgctag caccatgggc tgcgtgtgca gcagcaaccc cgagagatct
       61 gcgaattcag gtgctggtgc tggtgctggt gctatcctat ctagaatcct ctggcatgag
      121 atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa
      181 ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg
      241 aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc
      301 aggaagtaca tgaaatcagg gaatgtcaag gacctcctgc aagcctggga cctctattat
      361 catgtgttcc gacgaatctc aaagactagt tatccgtacg acgtaccaga ctacgcaccg
      421 gtcgccaccc tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      481 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      541 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      601 tggcccaccc tcgtgaccac cctgaccgcc ggcgtgcagt gcttcagccg ctaccccgac
      661 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      721 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      781 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      841 ctggggcaca agctggagta caactacatc agccacaacg tctatatcac cgcc
//
