LOCUS       sequence-147080-         663 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human codon optimized plasmid to express Cas9 t2a mCherry w/o NLS 
            and a sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 663)
  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 663)
  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: pBLO1811_Cas9_noNLS_human
FEATURES             Location/Qualifiers
     source          1..663
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(106..159)
                     /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"
ORIGIN
        1 ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa
       61 ctccttgatg atggccatgt tatcctcctc gcccttgctc accattgggc caggattctc
      121 ctcgacgtca ccgcatgtta gcagacttcc tctgccctct ccactgccgg attcggatcc
      181 gtcgcctccc agctgagaca ggtcgatccg tgtctcgtac aggccggtga tgctctggtg
      241 gatcagggtg gcgtccagca cctctttggt gctggtgtac ctcttccggt cgatggtggt
      301 gtcaaagtac ttgaaggcgg caggggctcc cagattggtc agggtaaaca ggtggatgat
      361 attctcggcc tgctctctga tgggcttatc ccggtgcttg ttgtaggcgg acagcacttt
      421 gtccagatta gcgtcggcca ggatcactct cttggagaac tcgctgatct gctcgatgat
      481 ctcgtccagg tagtgcttgt gctgttccac aaacagctgt ttctgctcat tatcctcggg
      541 ggagcccttc agcttctcat agtggctggc caggtacagg aagttcacat atttggaggg
      601 cagggccagt tcgtttccct tctgcagttc gccggcagag gccagcattc tcttccggcc
      661 gtt
//
