LOCUS       sequence-78108-e        1090 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NOTE: A new version of this plasmid is now available. See Addgene 
            plasmid 62988..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1090)
  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 1090)
  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: pSpCas9(BB)-2A-Puro (PX459)
FEATURES             Location/Qualifiers
     source          1..1090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(13..612)
                     /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     593..612
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(613..666)
                     /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(682..729)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 tagttagaat tctcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
       61 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgtgggg
      121 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
      181 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
      241 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
      301 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
      361 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
      421 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
      481 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
      541 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
      601 gtactcggtc attgggccag gattctcctc gacgtcaccg catgttagca gacttcctct
      661 gccctctcca ctgccgaatt cctttttctt ttttgcctgg ccggcctttt tcgtggccgc
      721 cggccttttg tcgcctccca gctgagacag gtcgatccgt gtctcgtaca ggccggtgat
      781 gctctggtgg atcagggtgg cgtccagcac ctctttggtg ctggtgtacc tcttccggtc
      841 gatggtggtg tcaaagtact tgaaggcggc aggggctccc agattggtca gggtaaacag
      901 gtggatgata ttctcggcct gctctctgat gggcttatcc cggtgcttgt tgtaggcgga
      961 cagcactttg tccagattag cgtcggccag gatcactctc ttggagaact cgctgatctg
     1021 ctcgatgatc tcgtccaggt agtgcttgtg ctgttccaca aacagctgtt tctgctcatt
     1081 atcctcgggg
//
