LOCUS       sequence-152416-        1070 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized S. pyogenes Cas9n (D10A nickase) 
            protein and blasticidin resistance from EFS promoter. Lentiviral 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1070)
  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 1070)
  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: lentiCas9n(D10A)-Blast
FEATURES             Location/Qualifiers
     source          1..1070
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(11..67)
                     /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(77..100)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(101..148)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 gcttggccat cggtccagga ttctcttcga catctccggc ttgtttcagc agagagaagt
       61 ttgttgcgcc ggatccctta tcgtcatcgt ctttgtaatc tttcttcttc ttagcctgtc
      121 cagccttctt tgtggcggca ggtcgcttgt cgcctcccag ctgagacagg tcgatccgtg
      181 tctcgtacag gccggtgatg ctctggtgga tcagggtggc gtccagcacc tctttggtgc
      241 tggtgtacct cttccggtcg atggtggtgt caaagtactt gaaggcggca ggggctccca
      301 gattggtcag ggtaaacagg tggatgatat tctcggcctg ctctctgatg ggcttatccc
      361 ggtgcttgtt gtaggcggac agcactttgt ccagattagc gtcggccagg atcactctct
      421 tggagaactc gctgatctgc tcgatgatct cgtccaggta gtgcttgtgc tgttccacaa
      481 acagctgttt ctgctcatta tcctcggggg agcccttcag cttctcatag tggctggcca
      541 ggtacaggaa gttcacatat ttggagggca gggccagttc gtttcccttc tgcagttcgc
      601 cggcagaggc cagcattctc ttccggccgt tttccagctc gaacagggag tacttaggca
      661 gcttgatgat caggtccttt ttcacttctt tgtagccctt ggcttccaga aagtcgatgg
      721 gattcttctc gaagctgctt ctttccatga tggtgatccc cagcagctct ttcacactct
      781 tcagtttctt ggacttgccc ttttccactt tggccaccac cagcacagaa taggccacgg
      841 tggggctgtc gaagccgccg tacttcttag ggtcccagtc cttctttctg gcgatcagct
      901 tatcgctgtt cctcttgggc aggatagact ctttgctgaa gccgcctgtc tgcacctcgg
      961 tctttttcac gatattcact tggggcatgc tcagcacttt ccgcacggtg gcaaaatccc
     1021 ggcccttatc ccacacgatc tccccggttt cgccgtttgt ctcgatcaga
//
