LOCUS       sequence-95589-e         649 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 649)
  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 649)
  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-5
FEATURES             Location/Qualifiers
     source          1..649
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        124..372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     124..144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     295..314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        391..466
                     /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 catagaggca
      121 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      181 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      241 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      301 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      361 ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      481 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      541 gacaaatggc tctagagtga gacccgttac ataacttacg gtaaatggcc cgcctggctg
      601 accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggg
//
