LOCUS       sequence-119272-         901 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses gRNA that targets stop codon for OCT4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 901)
  AUTHORS   Zhu Z, Verma N, Gonzalez F, Shi ZD, Huangfu D
  TITLE     A CRISPR/Cas-Mediated Selection-free Knockin Strategy in Human 
            Embryonic Stem Cells.
  JOURNAL   Stem Cell Reports. 2015 May 28. pii: S2213-6711(15)00128-9. doi: 
            10.1016/j.stemcr.2015.04.016.
  PUBMED    26028531
REFERENCE   2  (bases 1 to 901)
  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: "Stem Cell 
            Reports. 2015 May 28. pii: S2213-6711(15)00128-9. doi: 
            10.1016/j.stemcr.2015.04.016."
COMMENT     Sequence Label: Oct4-Cr1
FEATURES             Location/Qualifiers
     source          1..901
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     117..136
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        215..290
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(408..507)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(556..574)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(557..574)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(592..608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(592..608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(605..627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    616..632
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(640..646,647..664,665..670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    685..706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(791..813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
       61 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      121 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      181 aaggacgaaa caccgctccc atgcattcaa actggtttta gagctagaaa tagcaagtta
      241 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
      301 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
      361 tgcagacaaa tggctctaga ggtacgaatt caagggtggg cgcgccgacc cagctttctt
      421 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
      481 tatcagtcaa aataaaatca ttatttgcca tccagatccg agctcggtac caagcttgat
      541 gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
      601 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      661 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      721 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cctcgcgcgt ttcggtgatg
      781 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      841 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
      901 c
//
