LOCUS       sequence-167695-         895 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 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 895)
  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 895)
  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: pSpCas9(BB)-2A-Puro (PX459) V2.0
FEATURES             Location/Qualifiers
     source          1..895
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        127..375
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     127..147
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     298..317
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        394..469
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        566..851
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
       61 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      121 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
      181 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      241 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      301 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      361 aaaggacgaa acaccgggtc ttcgagaaga cctgttttag agctagaaat agcaagttaa
      421 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt
      481 agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct
      541 gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc ccgcctggct
      601 gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt
      661 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      721 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg
      781 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      841 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccc
//
