LOCUS       sequence-83341-e        1118 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid contains expression cassette for NmCas9 with N and C 
            NLS and an HA tag, a cassette for expression of tracrRNA, a cassette
            for cloning crRNA under the control of U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1118)
  AUTHORS   Hou Z, Zhang Y, Propson NE, Howden SE, Chu LF, Sontheimer EJ, 
            Thomson JA
  TITLE     Efficient genome engineering in human pluripotent stem cells using 
            Cas9 from Neisseria meningitidis.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 12.
  PUBMED    23940360
REFERENCE   2  (bases 1 to 1118)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Aug 12."
COMMENT     Sequence Label: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)
FEATURES             Location/Qualifiers
     source          1..1118
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     308..327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        392..640
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        640..732
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     promoter        774..792
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(811..829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        867..1115
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc
       61 caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc ggggactgtg
      121 ggcgatgtgc gctctgccca ctgacgggca ccggagcctc cgcgaagcag cgcaaaacgc
      181 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
      241 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
      301 aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc
      361 ccgatccgtc gacgtcaagg tcgggcagga agagggccta tttcccatga ttccttcata
      421 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
      481 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
      541 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
      601 tttcttggct ttatatatct tgtggaaagg acgaaacacc gaaatgagaa ccgttgctac
      661 aataaggccg tctgaaaaga tgtgccgcaa cgctctgccc cttaaagctt ctgctttaag
      721 gggcatcgtt tatttcggtt tttttgcggc cgcgtttaaa catcccgcga aattaatacg
      781 actcactata ggctgagttg gctgctgcca ccgctgagca ataactagca tatcgatatg
      841 gattcggatc taaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc
      901 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
      961 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     1021 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     1081 tggctttata tatcttgtgg aaaggacgaa acaccgga
//
