LOCUS       sequence-95575-e         665 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 nuclease and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 665)
  AUTHORS   Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T
  TITLE     Multiplex genome engineering in human cells using all-in-one 
            CRISPR/Cas9 vector system.
  JOURNAL   Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400.
  PUBMED    24954249
REFERENCE   2  (bases 1 to 665)
  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"; journalName: "Sci Rep."; 
            date: "2014-06-23"; volume: "4:5400. doi"; pages: " 
            10.1038/srep05400"
COMMENT     Sequence Label: pX330S-2
FEATURES             Location/Qualifiers
     source          1..665
                     /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"
ORIGIN
        1 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
       61 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcaggtct cactctagag
      121 gcatgtgagg 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 ggctctatga gacccgttac ataacttacg gtaaatggcc cgcctggctg
      601 accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg
      661 acgtc
//
