LOCUS       sequence-93128-e         877 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA, Puromycin 
            resistance, EFS Promoter driven.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 877)
  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 877)
  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: pL-CRISPR.EFS.PAC
FEATURES             Location/Qualifiers
     source          1..877
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(40..639)
                     /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(123..143)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     620..639
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(640..696)
                     /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(712..759)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 aattttgtaa tccagaggtt gattgtcgac ttaacgcgtt caggcaccgg gcttgcgggt
       61 catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag
      121 ccgctcgtag aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc
      181 gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg
      241 gtcgggcgag actccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc
      301 caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg
      361 cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc
      421 caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag
      481 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg
      541 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc
      601 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gggccggggt tctcctccac
      661 gtcgccggcc tgcttcagca ggctgaagtt ggtggcgccg ctgccgctag ctttcttttt
      721 cttagcttga ccagctttct tagtagcagc aggacgcttg tcgcctccca gctgagacag
      781 gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg atcagggtgg cgtccagcac
      841 ctctttggtg ctggtgtacc tcttccggtc gatggtg
//
