Difference between revisions of "Part:BBa K3454037"
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<partinfo>BBa_K3454037 short</partinfo> | <partinfo>BBa_K3454037 short</partinfo> | ||
− | This part is the template for AsCas12a extraction and purfication for reaction system. | + | This part is the template for AsCas12a extraction and purfication for reaction system.We got our wide-type AsCas12a protein from <bbpart>BBa_K2965021</bbpart>. 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=== | ===Characterization=== | ||
====Results==== | ====Results==== |
Latest revision as of 23:16, 27 October 2020
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:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 196
Illegal PstI site found at 3667
Illegal PstI site found at 3865 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3855
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 196
Illegal PstI site found at 3667
Illegal PstI site found at 3865 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1039
Illegal BsaI.rc site found at 2701
Illegal SapI.rc site found at 289