Coding

Part:BBa_K2933158

Designed by: Ruihan Dong   Group: iGEM19_TJUSLS_China   (2019-09-14)
Revision as of 08:04, 22 September 2019 by Xueqing (Talk | contribs)


His+Linker f+BlaB-14

This part encodes the fusion protein of His tag and BlaB-14 to promote the expression and purification of target protein(BlaB-14).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 51
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 647
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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 BlaB-14. It encodes a protein which is BlaB-14 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.

Molecular cloning

First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.We then extracted plasmids and performed double enzyme digestion verification.

Blab PCR.png
Figure 1. The PCR result of BlaB-14.

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Parameters
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