LOCUS       sequence-108082-         985 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 985)
  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 985)
  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..985
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(88..486)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(487..543)
                     /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(553..576)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(577..624)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
       61 attatcgata agcttgatat cgaattctta gccctcccac acataaccag agggcagcaa
      121 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
      181 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
      241 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
      301 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
      361 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
      421 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
      481 ggccatcggt ccaggattct cttcgacatc tccggcttgt ttcagcagag agaagtttgt
      541 tgcgccggat cccttatcgt catcgtcttt gtaatctttc ttcttcttag cctgtccagc
      601 cttctttgtg gcggcaggtc gcttgtcgcc tcccagctga gacaggtcga tccgtgtctc
      661 gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt
      721 gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt
      781 ggtcagggta aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg
      841 cttgttgtag gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga
      901 gaactcgctg atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag
      961 ctgtttctgc tcattatcct cgggg
//
