LOCUS       sequence-95573-e         522 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 522)
  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 522)
  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..522
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(164..239)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(258..506)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(316..335)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(486..506)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
       61 atgtaacggg tctcatagag ccatttgtct gcagaattgg cgcacgcgct aaaaacggac
      121 tagccttatt ttaacttgct atttctagct ctaaaacaaa aaagcaccga ctcggtgcca
      181 ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaaca
      241 ggtcttctcg aagacccggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg
      301 aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact
      361 gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg
      421 tttacagtca aattaattcc aattatctct ctaacagcct tgtatcgtat atgcaaatat
      481 gaaggaatca tgggaaatag gccctcacat gcctctagag tg
//
