LOCUS       sequence-162507-         915 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and a non-targeting sgRNA element from 
            U6 promoter. Lentiviral backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 915)
  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 915)
  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 - CTRL sgRNA
FEATURES             Location/Qualifiers
     source          1..915
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     133..152
                     /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 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
       61 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      121 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      181 ttatatatct tgtggaaagg acgaaacacc gaacctacgg gctacgatac ggttttagag
      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 tgatc
//
