LOCUS       sequence-78201-e        1087 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NOTE: A new version of this plasmid is now available. See Addgene 
            plasmid 62987.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1087)
  AUTHORS   Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F
  TITLE     Genome engineering using the CRISPR-Cas9 system.
  JOURNAL   Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. 
            Epub 2013 Oct 24.
  PUBMED    24157548
REFERENCE   2  (bases 1 to 1087)
  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"; doi: 
            "10.1038/nprot.2013.143"; journalName: "Nat Protoc"; date: 
            "2013-11"; volume: "8"; issue: "11"; pages: "2281-308"
COMMENT     Sequence Label: pSpCas9n(BB)-2A-Puro (PX462)
FEATURES             Location/Qualifiers
     source          1..1087
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(11..610)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     591..610
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(611..664)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(680..727)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 gttagaattc tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
       61 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgtggggcg
      121 cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
      181 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga
      241 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg
      301 ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg
      361 cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca
      421 ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct
      481 cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg
      541 cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
      601 actcggtcat tgggccagga ttctcctcga cgtcaccgca tgttagcaga cttcctctgc
      661 cctctccact gccgaattcc tttttctttt ttgcctggcc ggcctttttc gtggccgccg
      721 gccttttgtc gcctcccagc tgagacaggt cgatccgtgt ctcgtacagg ccggtgatgc
      781 tctggtggat cagggtggcg tccagcacct ctttggtgct ggtgtacctc ttccggtcga
      841 tggtggtgtc aaagtacttg aaggcggcag gggctcccag attggtcagg gtaaacaggt
      901 ggatgatatt ctcggcctgc tctctgatgg gcttatcccg gtgcttgttg taggcggaca
      961 gcactttgtc cagattagcg tcggccagga tcactctctt ggagaactcg ctgatctgct
     1021 cgatgatctc gtccaggtag tgcttgtgct gttccacaaa cagctgtttc tgctcattat
     1081 cctcggg
//
