Coding

Part:BBa_K3853003

Designed by: Gongyu Liu   Group: iGEM21_CPU_CHINA   (2021-09-20)


dCas9

For dCas9, two nuclease splicing active domains of CRISPR associated protein 9 (Cas9), RuvC and HNH, are mutated simultaneously (D10A & H840A; RuvC- & HNH-) to obtain a protein that has no nuclease activity. The dCas9 protein has no ability to cut DNA, but can still bind to specific DNA sequences under the single-gudie RNA. The codon of the protein gene sequence are optimized to increase its expression in E.coli. The introduction of this gene helps us immobilize the enzyme on dsDNA.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3576
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3960
    Illegal AgeI site found at 3304
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None