LOCUS       sequence-106834-         892 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9n (D10A nickase mutant) from S. pyogenes with 2A-Puro, and 
            cloning backbone for sgRNA (V2.0).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 892)
  AUTHORS   Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F
  TITLE     Genome engineering using the CRISPR-Cas9 system.
  JOURNAL   Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. 
            Epub 2013 Oct 24.
  PUBMED    24157548
REFERENCE   2  (bases 1 to 892)
  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/nprot.2013.143"; journalName: "Nat Protoc"; date: 
            "2013-11"; volume: "8"; issue: "11"; pages: "2281-308"
COMMENT     Sequence Label: pSpCas9n(BB)-2A-Puro (PX462) V2.0
FEATURES             Location/Qualifiers
     source          1..892
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        123..371
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     123..143
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     294..313
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        390..465
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        562..847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
      181 agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
      241 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
      301 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
      361 gacgaaacac cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata
      421 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      481 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      541 acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc
      601 gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg
      661 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      721 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca
      781 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      841 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc cc
//
