LOCUS       sequence-69388-e         788 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a S. pyogenes Cas9/dCas9 guide RNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 788)
  AUTHORS   Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein 
            LR, Thakore PI, Glass KA, Ousterout DG, Leong KW, Guilak F, Crawford
            GE, Reddy TE, Gersbach CA
  TITLE     RNA-guided gene activation by CRISPR-Cas9-based transcription 
            factors.
  JOURNAL   Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600.
  PUBMED    23892895
REFERENCE   2  (bases 1 to 788)
  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: "Nat 
            Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600."
COMMENT     Sequence Label: pSPgRNA
FEATURES             Location/Qualifiers
     source          1..788
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        220..295
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(329..349)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(329..347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(368..384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(368..384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(381..403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    392..408
                     /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(416..422,423..440,441..446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    461..482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(599..616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
       61 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
      121 atggactatc atatgcttat cgtaacttga aagtatttcg atttcttggc tttatatatc
      181 ttgtggaaag gacgaaacac cgggtcttcg agaagacctg ttttagagct agaaatagca
      241 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      301 ttccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      361 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      781 tccgcccc
//
