Composite

Part:BBa_K2933277

Designed by: Weisi Wang   Group: iGEM19_TJUSLS_China   (2019-09-15)
Revision as of 07:31, 24 September 2019 by Weisi (Talk | contribs) (→‎References)


RBS+Linker h+His+Linker f+BcII-194+T7 terminator

This part consists of RBS, protein coding sequence(His+Linker h+Sumo+Linker f+BcII-194) and T7 terminator,and the biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 93
    Illegal NheI site found at 907
  • 21
    COMPATIBLE WITH RFC[21]
  • 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 BcII. It encodes a protein which is BcII fused with His tag. The fusion protein is about 28.1 kD. It is convenient for us to purify our target protein.

Molecular cloning

First,we obtained BcII by PCR.

BcII-194 PCR.jpeg
Figure 1. The PCR result of BcII.

References

Sung-Kun Kim, Mara Demuth, Sara R. Schlesinger, Sung Joon Kim, Jonathan Urbanczyk, Robert W. Shaw & Hyunshun Shin (2016) Inhibition of Bacillusanthracis metallo-β- lactamase by compounds with hydroxamic acid functionality, Journal of Enzyme Inhibition and Medicinal Chemistry, 31:sup4, 132-137, DOI: 10.1080/14756366.2016.1222580

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