LOCUS       sequence-92597-e         550 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 550)
  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 550)
  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: pX330A-1x4
FEATURES             Location/Qualifiers
     source          1..550
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     53..72
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        149..224
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     465..487
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     479..496
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     480..496
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     506..525
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        506..524
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
       61 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      121 acgaaacacc gggtcttcga gaagacctgt tttagagcta gaaatagcaa gttaaaataa
      181 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gttttagagc
      241 tagaaatagc aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca attctgcaga
      301 caaatggctc tagagaccgg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      361 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacnccagc tggcgaaagg
      421 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      481 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      541 gccccccctc
//
