LOCUS       sequence-71903-e         856 bp    DNA     linear   UNA 14-APR-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 856)
  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 856)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..856
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     297..316
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        381..629
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     381..401
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     552..571
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        629..721
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     promoter        763..781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(800..818)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ggaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta
       61 ggcaccggtt caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg
      121 ctctgcccac tgacgggcac cggagcctcc gcgaagcagc gcaaaacgcc taaccctaag
      181 cagattcttc atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc
      241 gcactactca gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc
      301 atcagagcag attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg
      361 acgtcaaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac
      421 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
      481 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      541 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      601 tatatatctt gtggaaagga cgaaacaccg aaatgagaac cgttgctaca ataaggccgt
      661 ctgaaaagat gtgccgcaac gctctgcccc ttaaagcttc tgctttaagg ggcatcgttt
      721 atttcggttt ttttgcggcc gcgtttaaac atcccgcgaa attaatacga ctcactatag
      781 gctgagttgg ctgctgccac cgctgagcaa taactagcat atcgatatgg attcggatct
      841 aaggtcgggc aggaag
//
