LOCUS       sequence-128383-        1031 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized Cas9 protein and puromycin 
            resistance from EFS promoter and a DDX3Y 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 1031)
  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 1031)
  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 - DDX3Y sgRNA 2
FEATURES             Location/Qualifiers
     source          1..1031
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(322..342)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(349..372)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(401..612)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    681..798
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(846..921)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1001..1020)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tctgtggaag aagctgtcgt ccaccttggc catctcgttg ctgaagatct cttgcagata
       61 gcagatccgg ttcttccgtc tggtgtatct tcttctggcg gttctcttca gccgggtggc
      121 ctcggctgtt tcgccgctgt cgaacagcag ggctccgatc aggttcttct tgatgctgtg
      181 ccggtcggtg ttgcccagca ccttgaattt cttgctgggc accttgtact cgtcggtgat
      241 cacggcccag cccacagagt tggtgccgat gtccaggccg atgctgtact tcttgtcggc
      301 tgctgggact ccgtggatac cgaccttccg cttcttcttt ggggccatct tatcgtcatc
      361 gtctttgtaa tccatggtgg cggatcccgc gtcacgacac ctgtgttctg gcggcaaacc
      421 cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct
      481 cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc
      541 tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg
      601 tgcgctctgc ccactgacgg gcaccggagc cactcgagcc ggcgccaaag tggatctctg
      661 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
      721 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
      781 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctcaagat ctagaattca
      841 aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct tattttaact
      901 tgctatttct agctctaaaa ctggttttag ccccaacaag acggtgtttc gtcctttcca
      961 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
     1021 cattttaaaa c
//
