Coding

Part:BBa_K3044024

Designed by: Catharina Bang Jensen   Group: iGEM19_SDU-Denmark   (2019-10-10)
Revision as of 10:51, 14 October 2019 by Catharina (Talk | contribs)


Codon optimized Cas9 for expression in E. coli

Cas9 is a protein with endonuclease activity and induces a double stranded break in DNA.

Cas9 is the active protein in the CRISPR/Cas9 system II. It has its origin in Streptococcus Pyogenes and can be used as a gene editing tool for specific target genes.

Cas9-enzymes are DNA specific endonucleases, which perform double stranded breaks in the target DNA. To identify the target, the protein uses an RNA molecule referred to as a single guide RNA (sgRNA) which forms a complex with the Cas9 protein and functions as its guide towards the target. [1]

This Cas9 is made activated by making A10D and A840H mutations of BBa_K3044008 to activate the catalytic domains of the protein.


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 251
    Illegal BglII site found at 1325
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None