LOCUS       sequence-140717-         878 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 878)
  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 878)
  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 2
FEATURES             Location/Qualifiers
     source          1..878
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     121..140
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        219..294
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    342..459
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        528..739
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      759..768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             768..791
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             798..818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
       61 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
      121 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
      181 tggaaaggac gaaacaccgg ctcccttcaa gtgggatggt tttagagcta gaaatagcaa
      241 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      301 gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
      361 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
      421 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
      481 agagatccac tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac
      541 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
      601 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
      661 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
      721 gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc caccatggat tacaaagacg
      781 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
      841 ccgacaagaa gtacagcatc ggcctggaca tcggcacc
//
