LOCUS       sequence-148434-         700 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 700)
  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 700)
  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..700
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     103..122
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        381..666
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
       61 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
      121 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
      181 gggtcttcga gaagacctgt ttaagagcta tgctggaaac agcatagcaa gtttaaataa
      241 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gttttagagc
      301 tagaaatagc aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca attctgcaga
      361 caaatggctc tagaggtacc cgttacataa cttacggtaa atggcccgcc tggctgaccg
      421 cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt
      481 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      541 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag
      601 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      661 accatggtcg aggtgagccc cacgttctgc ttcactctcc
//
