LOCUS       sequence-83017-e        1015 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Replaces original lentiCRISPRv1 (Addgene Plasmid 49535) and produces
            ~10-fold higher titer virus. 3rd generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1015)
  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 1015)
  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: lentiCRISPR v2
FEATURES             Location/Qualifiers
     source          1..1015
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(56..652)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(139..159)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(653..709)
                     /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             complement(719..742)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(743..790)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgttcagg
       61 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
      121 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga
      181 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca
      241 gacccttgcc ctggtggtcg ggcgagactc cgacggtggc caggaaccac gcgggctcct
      301 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc
      361 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
      421 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
      481 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
      541 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
      601 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcggtccag
      661 gattctcttc gacatctccg gcttgtttca gcagagagaa gtttgttgcg ccggatccct
      721 tatcgtcatc gtctttgtaa tctttcttct tcttagcctg tccagccttc tttgtggcgg
      781 caggtcgctt gtcgcctccc agctgagaca ggtcgatccg tgtctcgtac aggccggtga
      841 tgctctggtg gatcagggtg gcgtccagca cctctttggt gctggtgtac ctcttccggt
      901 cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc cagattggtc agggtaaaca
      961 ggtggatgat attctcggcc tgctctctga tgggcttatc ccggtgcttg ttgta
//
