LOCUS       sequence-186625-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human RFK.
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: RFK gRNA (BRDN0001148883)
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        152..227
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    283..400
                     /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 cgtatcatgc ataccttcaa agttttagag ctagaaatag caagttaaaa
      181 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg
      241 gcgtaactag atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
      301 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      361 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      421 cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga taaagtttta
      481 aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg
      541 ctccggtgcc cgtcagtggg cagagcgcac
//
