LOCUS       sequence-93135-e         902 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 902)
  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 902)
  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..902
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(41..640)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(124..144)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     621..640
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(641..697)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(713..760)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 aaattttgta atccagaggt tgattgtcga cttaacgcgt tcaggcaccg ggcttgcggg
       61 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
      121 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
      181 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
      241 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
      301 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
      361 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
      421 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
      481 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
      541 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
      601 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggggccgggg ttctcctcca
      661 cgtcgccggc ctgcttcagc aggctgaagt tggtggcgcc gctgccgcta gctttctttt
      721 tcttagcttg accagctttc ttagtagcag caggacgctt gtcgcctccc agctgagaca
      781 ggtcgatccg tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca
      841 cctctttggt gctggtgtac ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg
      901 ca
//
