LOCUS       sequence-93134-e         822 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral Vector for SpCas9 Expression without sgRNA, Puromycin 
            resistance, EFS Promoter driven.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 822)
  AUTHORS   Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey 
            ME, Thielke A, Aster JC, Regev A, Ebert BL
  TITLE     Generation of mouse models of myeloid malignancy with combinatorial 
            genetic lesions using CRISPR-Cas9 genome editing.
  JOURNAL   Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951.
  PUBMED    24952903
REFERENCE   2  (bases 1 to 822)
  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: "Nat 
            Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951."
COMMENT     Sequence Label: pLKO5d.EFS.SpCas9.P2A.PAC
FEATURES             Location/Qualifiers
     source          1..822
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(311..331)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(338..361)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      361..370
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(390..601)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    670..787
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
       61 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
      121 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
      181 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
      241 ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcggct gctgggactc
      301 cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg tctttgtaat
      361 ccatggtggc ggatcccgcg tcacgacacc tgtgttctgg cggcaaaccc gttgcgaaaa
      421 agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg
      481 cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccg
      541 gttcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc
      601 cactgacggg caccggagcc actcgagccg gcgccaaagt ggatctctgc tgtccctgta
      661 ataaacccga aaattttgaa tttttgtaat ttgtttttgt aattctttag tttgtatgtc
      721 tgttgctatt atgtctacta ttctttcccc tgcactgtac cccccaatcc ccccttttct
      781 tttaaaattg tggatgaata ctgccatttg tctcaagatc ta
//
