LOCUS       sequence-187575-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human TWF2.
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: TWF2 gRNA (BRDN0001146396)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     58..77
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        157..232
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    288..405
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
       61 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      121 aaaggacgaa acaccgcctg agcattctgt gagtcggttt tagagctaga aatagcaagt
      181 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttaa
      241 gcttggcgta actagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa
      301 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
      361 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
      421 gacagcagag atccactttg gcgccggctc gagggggccc gggtgcaaag atggataaag
      481 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
      541 attggctccg gtgcccgtca gtgggcagag
//
