LOCUS       sequence-80471-e        1062 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Doxycycline-inducible lentiviral expression of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1062)
  AUTHORS   Wang T, Wei JJ, Sabatini DM, Lander ES
  TITLE     Genetic screens in human cells using the CRISPR-Cas9 system.
  JOURNAL   Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. 
            Epub 2013 Dec 12.
  PUBMED    24336569
REFERENCE   2  (bases 1 to 1062)
  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"; doi: 
            "10.1126/science.1246981"; journalName: "Science"; date: "2014-01-3-
            3"; volume: "343"; issue: "6166"; pages: "80-4"
COMMENT     Sequence Label: pCW-Cas9
FEATURES             Location/Qualifiers
     source          1..1062
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             52..648
                     /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(52..71)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     548..568
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             649..702
                     /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"
ORIGIN
        1 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agcaattcac catgaccgag
       61 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
      121 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
      181 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
      241 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
      301 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
      361 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
      421 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
      481 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
      541 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
      601 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccga aggtagaggt
      661 tctctcctca cttgtggtga tgttgaagaa aaccctggtc caatgtctag actggacaag
      721 agcaaagtca taaacggagc tctggaatta ctcaatggtg tcggtatcga aggcctgacg
      781 acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag
      841 aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca tcatacccac
      901 ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcatac
      961 cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca
     1021 gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tg
//
