LOCUS       sequence-85246-e         747 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  multiplex gRNA expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 747)
  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 747)
  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: pSQT1313
FEATURES             Location/Qualifiers
     source          1..747
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(161..180)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        complement(181..256)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(278..297)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     promoter        complement(298..546)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(356..375)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(526..546)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
       61 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtca
      121 tatgtagaga ggtacctcga gcggcccaag cttaaaaaaa ctgcctatac ggcagtgaac
      181 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
      241 tttctagctc taaaactgag acgcattaat cgtctcactg cctatacggc agtgaacggt
      301 gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt
      361 aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta
      421 ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattct
      481 aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag
      541 gccctcttcc tgcccgacct tggtaccgga tccagtcgac tgaattggtt cctttaaagc
      601 ctgctttttt gtacagctag cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
      661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      721 ctgataaatg cttcaataat attgaaa
//
