LOCUS       sequence-186349-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human RAF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 570)
  AUTHORS   Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, 
            Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, 
            Root DE
  TITLE     Optimized sgRNA design to maximize activity and minimize off-target 
            effects of CRISPR-Cas9.
  JOURNAL   Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
  PUBMED    26780180
REFERENCE   2  (bases 1 to 570)
  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. 2016 Jan 18. doi: 10.1038/nbt.3437."
COMMENT     Sequence Label: RAF1 gRNA (BRDN0001146172)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     54..73
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        153..228
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    284..401
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
       61 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
      121 gacgaaacac cggaccatgt ggacattagg tggttttaga gctagaaata gcaagttaaa
      181 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttaagctt
      241 ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg
      301 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
      361 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
      421 gcagagatcc actttggcgc cggctcgagg gggcccgggt gcaaagatgg ataaagtttt
      481 aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg
      541 gctccggtgc ccgtcagtgg gcagagcgca
//
