LOCUS       sequence-128324-        1018 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and a C16orf80 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 1018)
  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 1018)
  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 - C16orf80 sgRNA 1
FEATURES             Location/Qualifiers
     source          1..1018
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     131..150
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        230..305
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    353..470
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        539..750
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      770..779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             779..802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             809..829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
       61 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      121 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      181 atatatcttg tggaaaggac gaaacaccgt gtctgagaag taaacccgtg ttttagagct
      241 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      301 ggtgcttttt tgaattctag atcttgagac aaatggcagt attcatccac aattttaaaa
      361 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
      421 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
      481 acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg
      541 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
      601 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      661 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      721 tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga
      781 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
      841 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
      901 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      961 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggc
//
