LOCUS       sequence-83342-e        1060 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 1060)
  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 1060)
  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..1060
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(205..309)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(376..624)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(604..624)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     662..680
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(699..717)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        complement(759..851)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
ORIGIN
        1 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
       61 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gtttaaacaa aaaaatgaga
      361 cgtggaatcg tctccggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa
      421 atactttcaa gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc
      481 aaactaccca agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt
      541 tacagtcaaa ttaattctaa ttatctctct aacagccttg tatcgtatat gcaaatatga
      601 aggaatcatg ggaaataggc cctcttcctg cccgacctta gatccgaatc catatcgata
      661 tgctagttat tgctcagcgg tggcagcagc caactcagcc tatagtgagt cgtattaatt
      721 tcgcgggatg tttaaacgcg gccgcaaaaa aaccgaaata aacgatgccc cttaaagcag
      781 aagctttaag gggcagagcg ttgcggcaca tcttttcaga cggccttatt gtagcaacgg
      841 ttctcatttc ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact
      901 ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact
      961 acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag
     1021 tcaaattaat tctaattatc tctctaacag ccttgtatcg
//
