LOCUS       sequence-80977-e        1064 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 1064)
  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 1064)
  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 2
FEATURES             Location/Qualifiers
     source          1..1064
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(330..350)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(357..380)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      380..389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(409..620)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    689..806
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(854..929)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1008..1027)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc
      301 ttgtcggctg ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta
      361 tcgtcatcgt ctttgtaatc catggtggcg gatcccgcgt cacgacacct gtgttctggc
      421 ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat acggttctcc
      481 cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt ttaccccgcg
      541 ccaccttctc taggcaccgg ttcaattgcc gacccctccc cccaacttct cggggactgt
      601 gggcgatgtg cgctctgccc actgacgggc accggagcca ctcgagccgg cgccaaagtg
      661 gatctctgct gtccctgtaa taaacccgaa aattttgaat ttttgtaatt tgtttttgta
      721 attctttagt ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc
      781 ccccaatccc cccttttctt ttaaaattgt ggatgaatac tgccatttgt ctcaagatct
      841 agaattcaaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta
      901 ttttaacttg ctatttctag ctctaaaact ttacgtcgcc gtccagctcg gtgtttcgtc
      961 ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat
     1021 gatagtccat tttaaaacat aattttaaaa ctgcaaacta ccca
//
