LOCUS       sequence-162501-         952 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and an intergenic BCR/ABL-targeting 
            sgRNA element from U6 promoter. Lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 952)
  AUTHORS   Wang T, Birsoy K, Hughes NW, Krupczak KM, Post Y, Wei JJ, Lander ES,
            Sabatini DM
  TITLE     Identification and characterization of essential genes in the human 
            genome.
  JOURNAL   Science. 2015 Oct 15. pii: aac7041.
  PUBMED    26472758
REFERENCE   2  (bases 1 to 952)
  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: "Science. 
            2015 Oct 15. pii: aac7041."
COMMENT     Sequence Label: lentiCRISPR - Amplicon, BCR/ABL sgRNA 1
FEATURES             Location/Qualifiers
     source          1..952
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     123..142
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        221..296
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    344..461
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        530..741
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      761..770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             770..793
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             800..820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
       61 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      121 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      181 tgtggaaagg acgaaacacc ggatgacaga tgatggatgg gttttagagc tagaaatagc
      241 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      301 ttgaattcta gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg
      361 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
      421 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
      481 gcagagatcc actttggcgc cggctcgagt ggctccggtg cccgtcagtg ggcagagcgc
      541 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
      601 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
      661 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
      721 gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc gccaccatgg attacaaaga
      781 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
      841 agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
      901 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca ac
//
