LOCUS       sequence-93267-e         902 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor vector for genomic targeting of a Tetracycline-inducible Cas9 
            cassette to the human AAVS1/PPP1R12C locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 902)
  AUTHORS   Gonzalez F, Zhu Z, Shi ZD, Lelli K, Verma N, Li QV, Huangfu D
  TITLE     An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome 
            Editing in Human Pluripotent Stem Cells.
  JOURNAL   Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018.
  PUBMED    24931489
REFERENCE   2  (bases 1 to 902)
  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: "Cell Stem 
            Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018."
COMMENT     Sequence Label: Puro-Cas9 donor
FEATURES             Location/Qualifiers
     source          1..902
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..86
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(118..717)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(201..221)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     698..717
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(727..780)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    804..829
                     /label=SA
                     /note="splice acceptor site"
ORIGIN
        1 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
       61 ctggcaacta gaaggcacag tcgaggctga tcagcgggtt taaacgggcc ctctagatca
      121 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
      181 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag
      241 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
      301 cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc
      361 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
      421 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
      481 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
      541 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
      601 cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
      661 ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatctc
      721 gagcctaggg ccgggattct cctccacgtc accgcatgtt agaagacttc ctctgccctc
      781 tccgctgcca gatctctcga ggccctgtgg gaggaagaga agaggtcaga agcttgcccc
      841 actgtggggt ggaggggaca gataaaagta cccagaacca gagccacatt aaccggccct
      901 gg
//
