Coding
dCas 9

Part:BBa_K1051800:Design

Designed by: Shihong Chen   Group: iGEM13_Shenzhen_BGIC_ATCG   (2013-07-11)
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protein dCas9.Two mutations,D10A and H841A,from Cas9 gene without stop codon.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2740
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2886
    Illegal BsaI site found at 3740
    Illegal BsaI.rc site found at 1411


Design Notes

To mutation four Restriction Enzyme cutting sites(SpeI) and two codons(D10A & H841A),we design PCR primers to mutate them by PCR using Phution DNA polymerase.

Refrence http://www.ncbi.nlm.nih.gov/pubmed/?term=repurposing+CRISPR+as+an+RNA-Guided+Platfrom


Source

The Cas9 gene was got from Mr. Kai Tian,BGI Shenzhen China. The gene was splited into three parts to synthetize optimizing the codons in eucaryon.



References