LOCUS       sequence-98244-e         787 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 787)
  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 787)
  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_82
FEATURES             Location/Qualifiers
     source          1..787
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(37..56)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        complement(57..132)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(153..172)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        complement(173..248)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(269..288)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        complement(289..537)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(347..366)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(517..537)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        644..748
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tagagaggta cctcgagcgg cccaagctta aaaaaactgc ctatacggca gtgaacgcac
       61 cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc
      121 tagctctaaa accgaccaca tgaagcagca cgctgcctat acggcagtga acgcaccgac
      181 tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc
      241 tctaaaacgg acgtagcctt cgggcatgct gcctatacgg cagtgaacgg tgtttcgtcc
      301 tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg
      361 atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct
      421 acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc taattatctc
      481 tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcttc
      541 ctgcccgacc ttggtaccgg atccagtcga ctgaattggt tcctttaaag cctgcttttt
      601 tgtacagcta gcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      661 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      721 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      781 tcccttt
//
