LOCUS       Exported                 625 bp ds-DNA     linear   SYN 17-MAY-2021
DEFINITION  Cas9 from S. pyogenes with 2A-Puro, and cloning backbone for sgRNA 
            (V2.0).
ACCESSION   .
VERSION     .
KEYWORDS    pSpCas9(BB)-2A-Puro (PX459) V2.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 625)
  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 625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..625
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(330..350)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(357..422)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      422..431
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc
      301 ttgtcggctg ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta
      361 tcgtcatcgt ctttgtaatc aatatcatga tccttgtagt ctccgtcgtg gtccttatag
      421 tccatggtgg caccggtcca acctgaaaaa aagtgatttc aggcaggtgc tccaggtaat
      481 taaacattaa taccccacca accaaccatc ccttaaaccc ttacctcttg ctcagctaat
      541 tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc
      601 agagccgggg cgggcggcgc gaggc
//
