LOCUS       sequence-147072-         712 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 712)
  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 712)
  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..712
                     /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"
     CDS             complement(181..228)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga agcgcatgaa
       61 ctccttgatg atggccatgt tatcctcctc gcccttgctc accattgggc caggattctc
      121 ctcgacgtca ccgcatgtta gcagacttcc tctgccctct ccactgccgg attcggatcc
      181 ctttttcttt tttgcctggc cggccttttt cgtggccgcc ggccttttgt cgcctcccag
      241 ctgagacagg tcgatccgtg tctcgtacag gccggtgatg ctctggtgga tcagggtggc
      301 gtccagcacc tctttggtgc tggtgtacct cttccggtcg atggtggtgt caaagtactt
      361 gaaggcggca ggggctccca gattggtcag ggtaaacagg tggatgatat tctcggcctg
      421 ctctctgatg ggcttatccc ggtgcttgtt gtaggcggac agcactttgt ccagattagc
      481 gtcggccagg atcactctct tggagaactc gctgatctgc tcgatgatct cgtccaggta
      541 gtgcttgtgc tgttccacaa acagctgttt ctgctcatta tcctcggggg agcccttcag
      601 cttctcatag tggctggcca ggtacaggaa gttcacatat ttggagggca gggccagttc
      661 gtttcccttc tgcagttcgc cggcagaggc cagcattctc ttccggccgt tt
//
