Coding

Part:BBa_K3454037

Designed by: Ruijie Xiang   Group: iGEM20_ShanghaiTech_China   (2020-10-26)


AsCas12a

This part is the template for AsCas12a extraction and purfication for reaction system.We got our wide-type AsCas12a protein from BBa_K2965021. For higher expression yield in E. coli, we did codon optimization on the original coding sequence. Although the DNA sequences was changed, the amino acid sequence remained the same, so the protein was not changed after codon optimization.

Characterization

Results

We induced AsCas12a expression by IPTG and purified it by gravity-flow column with Ni-NTA Sefinose(TM) Resin. CBB(Coomassie brilliant blue) is used to characterize protein purification as shown in Figure 1.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 196
    Illegal PstI site found at 3667
    Illegal PstI site found at 3865
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 886
    Illegal PstI site found at 196
    Illegal PstI site found at 3667
    Illegal PstI site found at 3865
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3855
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 196
    Illegal PstI site found at 3667
    Illegal PstI site found at 3865
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 196
    Illegal PstI site found at 3667
    Illegal PstI site found at 3865
    Illegal NgoMIV site found at 3399
    Illegal AgeI site found at 2296
    Illegal AgeI site found at 2767
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1039
    Illegal BsaI.rc site found at 2701
    Illegal SapI.rc site found at 289


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