LOCUS       sequence-188283-         570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human NEK4.
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: NEK4 gRNA (BRDN0001146359)
FEATURES             Location/Qualifiers
     source          1..570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     59..78
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        158..233
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    289..406
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
       61 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
      121 gaaaggacga aacaccgctt ccctgtctcg acagcgagtt ttagagctag aaatagcaag
      181 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta
      241 agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
      301 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
      361 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
      421 ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa
      481 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
      541 aattggctcc ggtgcccgtc agtgggcaga
//
