LOCUS       sequence-80643-e         963 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and an EGFP targeting synthetic 
            single-guide RNA (sgRNA) element from U6 promoter. Lentiviral 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 963)
  AUTHORS   Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, 
            Heckl D, Ebert BL, Root DE, Doench JG, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout screening in human cells.
  JOURNAL   Science. 2014 Jan 3;343(6166):84-7. doi: 10.1126/science.1247005. 
            Epub 2013 Dec 12.
  PUBMED    24336571
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"; doi: 
            "10.1126/science.1247005"; journalName: "Science"; date: "2014-01-3-
            3"; volume: "343"; issue: "6166"; pages: "84-7"
COMMENT     Sequence Label: lentiCRISPR - EGFP sgRNA 6
FEATURES             Location/Qualifiers
     source          1..963
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     134..153
                     /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    355..472
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        541..752
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      772..781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             781..804
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             811..831
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 acgatacaag gctgttagag agataattag aattaatttg actgtaaaca caaagatatt
       61 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
      121 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
      181 tttatatatc ttgtggaaag gacgaaacac cggtgaaccg catcgagctg agttttagag
      241 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
      301 tcggtgcttt tttgaattct agatcttgag acaaatggca gtattcatcc acaattttaa
      361 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
      421 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
      481 ttacagggac agcagagatc cactttggcg ccggctcgag tggctccggt gcccgtcagt
      541 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
      601 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      661 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      721 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg
      781 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      841 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
      901 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
      961 aac
//
