LOCUS       sequence-128367-         988 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and a C9orf114 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 988)
  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 988)
  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 - C9orf114 sgRNA 2
FEATURES             Location/Qualifiers
     source          1..988
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     125..144
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        224..299
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    347..464
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        533..744
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      764..773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             773..796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             803..823
                     /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 ccgcaggcgg gctcacctcc gtggttttag agctagaaat
      241 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      301 tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      361 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      421 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      481 acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca gtgggcagag
      541 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      601 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      661 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      721 aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca tggattacaa
      781 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      841 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      901 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      961 ccggcacagc atcaagaaga acctgatc
//
