LOCUS       sequence-83038-e         972 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and puromycin resistance from EF-1a promoter.
            3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 972)
  AUTHORS   Sanjana NE, Shalem O, Zhang F
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 972)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2014-08"; volume: "11"; issue: 
            "8"; pages: "783-4"
COMMENT     Sequence Label: lentiGuide-Puro
FEATURES             Location/Qualifiers
     source          1..972
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             36..635
                     /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(36..55)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     532..552
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(704..724)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac cgagtacaag cccacggtgc
       61 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
      121 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
      181 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
      241 acggcgccgc cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
      301 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
      361 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
      421 gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
      481 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
      541 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
      601 gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac aatcaacctc
      661 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
      721 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
      781 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
      841 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
      901 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
      961 cggaactcat cg
//
