LOCUS       sequence-128395-         952 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 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 - AAVS1 sgRNA
FEATURES             Location/Qualifiers
     source          1..952
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     125..144
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        223..298
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    346..463
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        532..743
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      763..772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             772..795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             802..822
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
       61 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      121 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      181 cttgtggaaa ggacgaaaca ccggggccac tagggacagg atgttttaga gctagaaata
      241 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      301 ttttgaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
      361 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
      421 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
      481 cagcagagat ccactttggc gccggctcga gtggctccgg tgcccgtcag tgggcagagc
      541 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      601 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      661 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      721 acgggtttgc cgccagaaca caggtgtcgt gacgcgggat ccgccaccat ggattacaaa
      781 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
      841 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
      901 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg ca
//
