LOCUS       sequence-447375-        1389 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses BirA-FRB-NLS fusion from ADH1 promoter; yeast BioID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1389)
  AUTHORS   Pfannenstein J, Tyryshkin M, Gulden ME, Doud EH, Mosley AL, Reese JC
  TITLE     Characterization of BioID tagging systems in budding yeast and 
            exploring the interactome of the Ccr4-Not complex.
  JOURNAL   G3 (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221.
  PUBMED    39271111
REFERENCE   2  (bases 1 to 1389)
  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: "G3 
            (Bethesda). 2024 Sep 13:jkae221. doi: 10.1093/g3journal/jkae221."
COMMENT     Sequence Label: pRS416-ADH1p-BirA-FRB-NLS-HA3
FEATURES             Location/Qualifiers
     source          1..1389
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..354,355..357,358..966)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MSKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLR
                     KLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQ
                     GWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLII
                     GDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             985..1263
                     /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             1273..1293
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1294..1383
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
ORIGIN
        1 atgtccaagg acaacaccgt gcccctgaag ctgatcgccc tgctggccaa cggcgagttc
       61 cactctggcg agcagctggg agagaccctg ggaatgagca gagccgccat caacaagcac
      121 atccagacac tgagagactg gggagtggac gtgttcaccg tgcctggcaa gggctacagc
      181 ctgcctgagc ctatccagct gctgaacgcc aagcagatcc tgggacagct ggatggcgga
      241 agcgtggccg tgctgcctgt gatcgactcc accaatcagt acctgctgga cagaatcgga
      301 gagctgaagt ccggcgacgc ctgcatcgcc gagtaccagc aggctggcag aggaggcaga
      361 ggacggaagt ggttcagccc attcggagcc aacctgtacc tgtccatgtt ctggagactg
      421 gagcagggac ctgctgctgc catcggactg agtctggtga tcggaatcgt gatggccgag
      481 gtgctgagaa agctgggagc cgacaaggtg agagtgaagt ggcctaatga cctgtacctc
      541 caggaccgca agctggctgg catcctggtg gagctgacag gcaagacagg cgatgccgct
      601 cagatcgtga tcggagccgg aatcaacatg gccatgagaa gagtggagga gagcgtggtg
      661 aaccagggct ggatcaccct gcaggaggct ggcatcaacc tggaccggaa caccctggcc
      721 gccatgctga tcagagagct gagagccgct ctggagctgt tcgagcagga gggactggct
      781 ccttacctga gcagatggga gaagctggac aacttcatca acagacctgt gaagctgatc
      841 atcggcgaca aggaaatctt cggcatctcc agaggaatcg acaagcaggg agctctgctg
      901 ctggagcagg acggaatcat caagccctgg atgggcggag aaatctccct gagaagcgca
      961 gagaagggtt ctggtggtgc tagcatcctc tggcatgaga tgtggcatga aggcctggaa
     1021 gaggcatctc gtttgtactt tggggaaagg aacgtgaaag gcatgtttga ggtgctggag
     1081 cccttgcatg ctatgatgga acggggcccc cagactctga aggaaacatc ctttaatcag
     1141 gcctatggtc gagatttaat ggaggcccaa gagtggtgca ggaagtacat gaaatcaggg
     1201 aatgtcaagg acctcctcca agcctgggac ctctattatc atgtgttccg acgaatctca
     1261 aagctcgagc caccaaaaaa gaaaagaaaa gtttacccat acgatgttcc tgactatgcg
     1321 ggctatccgt atgacgtccc ggactatgca ggatcttatc catatgacgt tccagattac
     1381 gctgcttga
//
