LOCUS       sequence-98235-e         858 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses a pair of control multiplex gRNAs targeting EGFP for use 
            with dimeric hFokI-dCas9 nuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 858)
  AUTHORS   Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin
            MJ, Aryee MJ, Joung JK
  TITLE     Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome 
            editing.
  JOURNAL   Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908.
  PUBMED    24770325
REFERENCE   2  (bases 1 to 858)
  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 
            Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908."
COMMENT     Sequence Label: RFN_EGFP_47
FEATURES             Location/Qualifiers
     source          1..858
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    15..34
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        55..130
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    131..150
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        171..246
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    247..266
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(450..469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaggacgaaa caccgttcac tgccgtatag gcagaagttc agcgtgtccg gcgagtttta
       61 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
      121 gagtcggtgc gttcactgcc gtataggcag ccgtccagct cgaccaggat gttttagagc
      181 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      241 cggtgcgttc actgccgtat aggcagtttt tttaagcttg ggccgctcga ggtacctctc
      301 tacatatgac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      841 gccagttacc ttcggaaa
//
