LOCUS       sequence-186471-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human BUB1.
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: BUB1 gRNA (BRDN0001145062)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     55..74
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        154..229
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    285..402
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
       61 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      121 ggacgaaaca ccgtgatgaa tcttgggtca ttggttttag agctagaaat agcaagttaa
      181 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttaagct
      241 tggcgtaact agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
      301 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
      361 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
      421 agcagagatc cactttggcg ccggctcgag ggggcccggg tgcaaagatg gataaagttt
      481 taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt
      541 ggctccggtg cccgtcagtg ggcagagcgc
//
