LOCUS       sequence-98972-e         952 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor plasmid for HR-mediated recombination to insert puromycin 
            resistance and append a destabilization domain to the N-terminus of 
            TCOF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 952)
  AUTHORS   Park A, Won ST, Pentecost M, Bartkowski W, Lee B
  TITLE     CRISPR/Cas9 allows efficient and complete knock-in of a 
            destabilization domain-tagged essential protein in a human cell 
            line, allowing rapid knockdown of protein function.
  JOURNAL   PLoS One. 2014 Apr 17;9(4):e95101. doi: 
            10.1371/journal.pone.0095101. eCollection 2014.
  PUBMED    24743236
REFERENCE   2  (bases 1 to 952)
  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: "PLoS One.";
            date: "2014-04-17"; volume: "9(4):e95101. doi"; pages: " 
            10.1371/journal.pone.0095101. eCollection 2014"
COMMENT     Sequence Label: PuroR-P2A-DD-TCOF1-donor
FEATURES             Location/Qualifiers
     source          1..952
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             71..127
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(128..130,131..451)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
ORIGIN
        1 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
       61 cggatccgga gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc
      121 cggtcccatg ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa
      181 gcgcggccag acctgtgtgg tgcactacac cgggatgctt gaagatggaa agaaagtcga
      241 ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg
      301 aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc
      361 tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct
      421 cgtcttcgat gtggagcttc taaaaccgga aggtggaggt ggatctggtg gaggtggatc
      481 tggtggcggc ggttcaatgg ccgaggccag gaagcggcgg gagctacttc ccctgatcta
      541 ccaccatctg ctgcgggctg gctatgtgcg tgcggcgcgg gaagtgaagg agcagagcgg
      601 ccaggtaagc gttcgtgggc cgtgtgcgag ggccgcgtgc aagatgtgga gatcagcggc
      661 ccgcggcccg cgccccgtcc ccaggcgacc cggcaggcgc ccggagccgg gtcccgcagt
      721 gctcgacggc gcggccaggg gtaccggagg agccgcaatc tctgccttcc cactgcgact
      781 tcagttcccc tgggcctcac tttcctcatc cgggcagtgg gttgggccag aaagtgtgct
      841 cgcaggggcc gaactttgta attcccgctt cccttacctc cacgcccttc ctggtagcgg
      901 gtattttaag tttcctaagt ctcccgccac gtggctaggc tctgcgcggc cc
//
