LOCUS       sequence-107800-         977 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NOTE: A new version of this plasmid is now available. See Addgene 
            plasmid 62987.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 977)
  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 977)
  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)
FEATURES             Location/Qualifiers
     source          1..977
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        131..379
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     131..151
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     302..321
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        398..473
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        570..855
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
       61 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      121 gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
      181 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
      241 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
      301 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
      361 gtggaaagga cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag
      421 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      481 ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa
      541 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
      601 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
      661 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      721 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      781 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      841 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      901 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      961 gcgggggggg ggggggg
//
