LOCUS       sequence-146922-         692 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 692)
  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 692)
  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..692
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     116..135
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        394..679
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
       61 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
      121 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
      181 aggacgaaac accgggtctt cgagaagacc tgtttaagag ctatgctgga aacagcatag
      241 caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
      301 tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg
      361 ccaattctgc agacaaatgg ctctagaggt acccgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac
      481 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      541 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      601 gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      661 agtcatcgct attaccatgg tcgaggtgag cc
//
