LOCUS       sequence-128396-        1056 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and an AAVS1-targeting sgRNA element 
            from U6 promoter. Lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1056)
  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 1056)
  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 - AAVS1 sgRNA
FEATURES             Location/Qualifiers
     source          1..1056
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(323..343)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(350..373)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(402..613)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    682..799
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(847..922)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1001..1020)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gtctgtggaa gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc tcttgcagat
       61 agcagatccg gttcttccgt ctggtgtatc ttcttctggc ggttctcttc agccgggtgg
      121 cctcggctgt ttcgccgctg tcgaacagca gggctccgat caggttcttc ttgatgctgt
      181 gccggtcggt gttgcccagc accttgaatt tcttgctggg caccttgtac tcgtcggtga
      241 tcacggccca gcccacagag ttggtgccga tgtccaggcc gatgctgtac ttcttgtcgg
      301 ctgctgggac tccgtggata ccgaccttcc gcttcttctt tggggccatc ttatcgtcat
      361 cgtctttgta atccatggtg gcggatcccg cgtcacgaca cctgtgttct ggcggcaaac
      421 ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc
      481 tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt
      541 ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat
      601 gtgcgctctg cccactgacg ggcaccggag ccactcgagc cggcgccaaa gtggatctct
      661 gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt
      721 agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat
      781 cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtctcaaga tctagaattc
      841 aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
      901 ttgctatttc tagctctaaa acatcctgtc cctagtggcc ccggtgtttc gtcctttcca
      961 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
     1021 cattttaaaa cataatttta aaactgcaaa ctaccc
//
