LOCUS       sequence-156858-         963 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human PIM2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 963)
  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 963)
  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: PIM2 gRNA (BRDN0001149335)
FEATURES             Location/Qualifiers
     source          1..963
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     133..152
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        232..307
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    363..480
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        638..849
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
       61 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      121 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      181 ttatatatct tgtggaaagg acgaaacacc gatcctgata gacctacgcc ggttttagag
      241 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
      301 tcggtgcttt tttaagcttg gcgtaactag atcttgagac aaatggcagt attcatccac
      361 aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
      421 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
      481 cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg
      541 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc
      601 ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
      661 gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc
      721 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
      781 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
      841 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
      901 ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt
      961 cgg
//
