LOCUS       sequence-97646-e        2399 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor plasmid for HR-mediated recombination to insert blasticidin 
            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 2399)
  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 2399)
  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: BlastR-P2A-DD-TCOF1-donor
FEATURES             Location/Qualifiers
     source          1..2399
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             759..1154
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1164..1220
                     /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(1221..1223,1224..1544)
                     /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 ggatcccagc aatttcgggt gtctttatgc ctcagcttag atatctctct ggggttcggt
       61 gctcctcctt ggggcgccca taggctgcat ctgtccctga caaggcacac aagatattga
      121 actatagttg cttggttacg gacagtcgtc tctgtgagct ttgagggcaa ctgccatgta
      181 ttatgcatct tttgaatccc cctagcctag ccgggtgcta gcccaagtta ggtgcttatt
      241 acagatttgt tcagttagcg aatgaatcaa tgaataaact ggcaattcgt gtctggttca
      301 tgggcaccaa aaaccgtgcg aaatggataa atgagcaggt aatggcagta caaagttcgg
      361 atctagagaa agatccagat tagggagttc agagatgagt aaaacgcaga cccagccatc
      421 agagaattaa caaggaagat aaatctcttc aagaagccag ccggaaggat taaaagagag
      481 tcgtccaagt cccggccggc cccgggggcg ggtccctgtc tagccccgca tgctccgcgc
      541 caatgggcgg tattgttgat cacgagtctc cgcccccttc gccttgaggg gcggggccgg
      601 ccactcccgg agaggggact acgtttcccg gcgcgccgcg cggccggagc gaaagaggag
      661 ccggaagtgg ggcgcgcgag gtctaagggc gcgagggaag tggcgggcgg ggactaaggc
      721 ggggcgtgca ggtagccggc cggccggggg tcgcgggtat ggccaagcct ttgtctcaag
      781 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
      841 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
      901 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
      961 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     1021 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     1081 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     1141 atgtgtggga gggcggatcc ggagccacga acttctctct gttaaagcaa gcaggagacg
     1201 tggaagaaaa ccccggtccc atgggagtgc aggtggaaac catctcccca ggagacgggc
     1261 gcaccttccc caagcgcggc cagacctgtg tggtgcacta caccgggatg cttgaagatg
     1321 gaaagaaagt cgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     1381 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     1441 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     1501 cacatgccac tctcgtcttc gatgtggagc ttctaaaacc ggaaggtgga ggtggatctg
     1561 gtggaggtgg atctggtggc ggcggttcaa tggccgaggc caggaagcgg cgggagctac
     1621 ttcccctgat ctaccaccat ctgctgcggg ctggctatgt gcgtgcggcg cgggaagtga
     1681 aggagcagag cggccaggta agcgttcgtg ggccgtgtgc gagggccgcg tgcaagatgt
     1741 ggagatcagc ggcccgcggc ccgcgccccg tccccaggcg acccggcagg cgcccggagc
     1801 cgggtcccgc agtgctcgac ggcgcggcca ggggtaccgg aggagccgca atctctgcct
     1861 tcccactgcg acttcagttc ccctgggcct cactttcctc atccgggcag tgggttgggc
     1921 cagaaagtgt gctcgcaggg gccgaacttt gtaattcccg cttcccttac ctccacgccc
     1981 ttcctggtag cgggtatttt aagtttccta agtctcccgc cacgtggcta ggctctgcgc
     2041 ggccccccct ggggcaagga ggttgctgcg agtctcgggg gtggattgcg accagcccct
     2101 cgggggagcg cctggaaaag ggaccctaca gcctcttagt tgagctggaa ctcaggcctt
     2161 tagcgattgt ctgtactgag acggtcagct ttggtactcg aactcgtggt acagtcattt
     2221 cttacttgat taattcaact gttaatccgg tactcagggt gcttcctggt gttcgaatgg
     2281 tgagcaccat ctctcagccc tcctggagct tacactctag taaggtagac aaatcaaaga
     2341 atcacaaaag tgactatatg cttagcaccg caataagggc cggccaaaag ggcgaattc
//
