LOCUS       sequence-185405-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human NEK2.
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: NEK2 gRNA (BRDN0001146193)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     56..75
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        155..230
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    286..403
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
       61 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
      121 aggacgaaac accggccgct gccagaagat ccgggtttta gagctagaaa tagcaagtta
      181 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttaagc
      241 ttggcgtaac tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
      301 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
      361 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
      421 cagcagagat ccactttggc gccggctcga gggggcccgg gtgcaaagat ggataaagtt
      481 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      541 tggctccggt gcccgtcagt gggcagagcg
//
