LOCUS       sequence-71688-e         925 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one plasmid targeting human OCT4, cuts roughly ~84bp 
            downstream of stop codon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 925)
  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 925)
  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-crRNA(OCT4)-NLS-NmCas9-HA-NLS(s)
FEATURES             Location/Qualifiers
     source          1..925
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(198..302)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(397..645)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(455..474)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(625..645)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     683..701
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(720..738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        complement(780..872)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
ORIGIN
        1 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
       61 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      121 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
      181 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
      241 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
      301 cgcacatttc cccgaaaagt gccacctgac gtcgtttaaa caaaaaaacg aaatgagaaa
      361 gggagctaca acaaaccctg gcacaaactc caggtcggtg tttcgtcctt tccacaagat
      421 atataaagcc aagaaatcga aatactttca agttacggta agcatatgat agtccatttt
      481 aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt
      541 ttgtactaat atctttgtgt ttacagtcaa attaattcta attatctctc taacagcctt
      601 gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcttcct gcccgacctt
      661 agatccgaat ccatatcgat atgctagtta ttgctcagcg gtggcagcag ccaactcagc
      721 ctatagtgag tcgtattaat ttcgcgggat gtttaaacgc ggccgcaaaa aaaccgaaat
      781 aaacgatgcc ccttaaagca gaagctttaa ggggcagagc gttgcggcac atcttttcag
      841 acggccttat tgtagcaacg gttctcattt cggtgtttcg tcctttccac aagatatata
      901 aagccaagaa atcgaaatac tttca
//
