Coding
Part:BBa_K2933155
Designed by: Ruihan Dong Group: iGEM19_TJUSLS_China (2019-09-14)
His+Linker f+VIM-66
This part encodes the fusion protein of His tag and VIM-66 to promote the expression and purification of target protein(VIM-66).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 51
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 824
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.
Molecular cloning
We insert VIM-66 gene into the standard vector then transfer it into E.coli.
Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification
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Categories
Parameters
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