Coding

Part:BBa_K4335013

Designed by: Hang Zhang   Group: iGEM22_UESTC-BioTech   (2022-10-10)


SpCas9

The Cas9 protein is part of the bacteria's CRISPR/Cas9 immune defense mechanism to identify and destroy foreign DNA. By incorporating the foreign DNA into the bacteria's own DNA, it has a memory of any prior foreign DNA that the bacteria has encountered.

The Cas9 endonuclease can be used either with the tracrRNA and crRNA (CRISPR RNA- Clustered Regularly Interspaced Short Palindromic Repeats) or with a guide RNA to cleave foreign DNA at a species locations. This makes exceptionally useful for DNA cloning in synthetic biology.

Usage

We found a SpCas9 from iGEM16_Cambridge-JIC in the previous part [BBa_K2148013] .

This part is based on the original Cas9 part in the registry, BBa_K1218011. To enhance the expression of Cas9 in the chloroplast chassis, Cambridge-JIC have codon-optimised it using the codon-optimisation software developed by Saul Purton.

The amino acid and nucleotide sequences of BBa_K2148013 and BBa_K2148013 were compared.

The comparison of nucleotide sequences of BBa_K2148013 and BBa_K4335013 showed that the matching rate was 67.96%.

The amino acid sequences of BBa_K2148013 and BBa_K4335013 were compared and the matching degree was 100%.

Result

See the part [BBa_K4335020] for result.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 241
    Illegal PstI site found at 787
    Illegal PstI site found at 2209
    Illegal PstI site found at 2413
    Illegal PstI site found at 2443
    Illegal PstI site found at 3655
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 241
    Illegal PstI site found at 787
    Illegal PstI site found at 2209
    Illegal PstI site found at 2413
    Illegal PstI site found at 2443
    Illegal PstI site found at 3655
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 248
    Illegal BglII site found at 1043
    Illegal BglII site found at 1733
    Illegal BglII site found at 3812
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 241
    Illegal PstI site found at 787
    Illegal PstI site found at 2209
    Illegal PstI site found at 2413
    Illegal PstI site found at 2443
    Illegal PstI site found at 3655
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 241
    Illegal PstI site found at 787
    Illegal PstI site found at 2209
    Illegal PstI site found at 2413
    Illegal PstI site found at 2443
    Illegal PstI site found at 3655
    Illegal NgoMIV site found at 1075
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 3468


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