LOCUS       sequence-151234-         861 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 861)
  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 861)
  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-1x2
FEATURES             Location/Qualifiers
     source          1..861
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        120..368
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     120..140
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     291..310
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        387..462
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     703..725
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     717..734
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     718..734
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     744..763
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        744..762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(810..830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(810..828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
       61 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg
      121 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      181 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      241 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      301 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
      361 gaaacaccgg gtcttcgaga agacctgttt tagagctaga aatagcaagt taaaataagg
      421 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt tttagagcta
      481 gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca
      541 aatggctcta gagaccggcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      601 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      661 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      721 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
      781 cccccctcga ggtcctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
      841 taatcatggt catagctgtt t
//
