LOCUS       sequence-150395-         574 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human codon optimized plasmid to express Cas9 t2a mCherry w/2xNLS 
            and a sgRNA .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 574)
  AUTHORS   Oakes BL, Nadler DC, Flamholz A, Fellmann C, Staahl BT, Doudna JA, 
            Savage DF
  TITLE     Profiling of engineering hotspots identifies an allosteric 
            CRISPR-Cas9 switch.
  JOURNAL   Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3528.
  PUBMED    27136077
REFERENCE   2  (bases 1 to 574)
  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. 2016 May 2. doi: 10.1038/nbt.3528."
COMMENT     Sequence Label: pBLO1806_Cas9_2xNLS_human
FEATURES             Location/Qualifiers
     source          1..574
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(314..334)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(341..406)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      406..415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agaagctgtc gtccaccttg gccatctcgt tgctgaagat ctcttgcaga tagcagatcc
       61 ggttcttccg tctggtgtat cttcttctgg cggttctctt cagccgggtg gcctcggctg
      121 tttcgccgct gtcgaacagc agggctccga tcaggttctt cttgatgctg tgccggtcgg
      181 tgttgcccag caccttgaat ttcttgctgg gcaccttgta ctcgtcggtg atcacggccc
      241 agcccacaga gttggtgccg atgtccaggc cgatgctgta cttcttgtcg gctgctggga
      301 ctccgtggat accgaccttc cgcttcttct ttggggccat cttatcgtca tcgtctttgt
      361 aatcaatatc atgatccttg tagtctccgt cgtggtcctt atagtccatg gtggcaccgg
      421 tccaacctga aaaaaagtga tttcaggcag gtgctccagg taattaaaca ttaatacccc
      481 accaaccaac catcccttaa acccttacct cttgctcagc taattacagc ccggaggaga
      541 agggccgtcc cgcccgctca cctgtgggag taac
//
