Difference between revisions of "Part:BBa K3853003"
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− | + | 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 codons of the protein gene sequence are optimized to increase its expression in <i>E.coli</i>. | |
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Revision as of 21:30, 7 October 2021
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 codons of the protein gene sequence are optimized to increase its expression in E.coli.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3576
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3960
Illegal AgeI site found at 3304 - 1000COMPATIBLE WITH RFC[1000]