LOCUS       sequence-141733-         798 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 798)
  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 798)
  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..798
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    74..95
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(110..115,116..133,134..140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    148..164
                     /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)."
     primer_bind     153..175
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     172..188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     172..188
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     207..227
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        209..227
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        complement(261..336)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(355..603)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(583..603)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(617..636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(618..636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(646..663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(646..662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(655..677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
       61 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      121 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      181 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
      241 agctccaccg cggaaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact
      301 agccttattt taacttgcta tttctagctc taaaacaggt cttctcgaag acccggtgtt
      361 tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacgataag
      421 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta
      481 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat
      541 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
      601 ctcggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
      661 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
      721 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
      781 gcgcagcctg aatggcga
//
