LOCUS       sequence-202875-         943 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 upon induction with doxicycline..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 943)
  AUTHORS   Castano J, Bueno C, Jimenez-Delgado S, Roca-Ho H, Fraga MF, 
            Fernandez AF, Nakanishi M, Torres-Ruiz R, Rodriguez-Perales S, 
            Menendez P
  TITLE     Generation and characterization of a human iPSC cell line expressing
            inducible Cas9 in the ""safe harbor"" AAVS1 locus.
  JOURNAL   Stem Cell Res. 2017 May;21:137-140. doi: 10.1016/j.scr.2017.04.011. 
            Epub 2017 Apr 22.
  PUBMED    28677529
REFERENCE   2  (bases 1 to 943)
  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: "Stem Cell 
            Res."; date: "2017-05"; volume: "21:137-140. doi"; pages: " 
            10.1016/j.scr.2017.04.011. Epub 2017 Apr 22"
COMMENT     Sequence Label: pAAVS1-TRE-Cas9- puro-polyA-CAG-rtTA
FEATURES             Location/Qualifiers
     source          1..943
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(24..623)
                     /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(107..127)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     604..623
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(633..686)
                     /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    710..735
                     /label=SA
                     /note="splice acceptor site"
ORIGIN
        1 gggcattgtt acggggctct agatcaggca ccgggcttgc gggtcatgca ccaggtgcgc
       61 ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg
      121 aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc
      181 actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg
      241 acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc
      301 tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg
      361 aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg
      421 tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc
      481 tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg
      541 gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc
      601 accgtgggct tgtactcggt catctcgagc ctagggccgg gattctcctc cacgtcaccg
      661 catgttagaa gacttcctct gccctctccg ctgccagatc tctcgaggcc ctgtgggagg
      721 aagagaagag gtcagaagct tgccccactg tggggtggag gggacagata aaagtaccca
      781 gaaccagagc cacattaacc ggccctggga atataaggtg ggtcccagct cggggacaca
      841 ggatccctgg aggcagcaaa catgctgtcc tgaagtgtac ataagggccc gggtttggaa
      901 ggaataaact agcttgagct ctcggacccc ctggaagatg gcc
//
