LOCUS       sequence-184375-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human EGFR.
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: EGFR gRNA (BRDN0001149464)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     62..81
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        161..236
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    292..409
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
       61 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
      121 gtggaaagga cgaaacaccg tgtcaccaca taattacctg gttttagagc tagaaatagc
      181 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      241 ttaagcttgg cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag
      301 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
      361 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
      421 cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat
      481 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
      541 gggaattggc tccggtgccc gtcagtgggc
//
