Composite

Part:BBa_K2933178

Designed by: Weisi Wang   Group: iGEM19_TJUSLS_China   (2019-09-14)
Revision as of 12:51, 21 September 2019 by Weisi (Talk | contribs)


Tac promoter+RBS a+Linker g+GST+Linker e+BlaB-14

This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+BlaB-14),and the biological module can be built into E.coli for protein expression.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1355
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 181


Usage and Biology


Molecular cloning

First, we used the vector pGEX-6p-1 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-14-PCR.png
Figure 1. (a) The PCR result of BlaB-14. (b) The verification results by enzyme digestion.

After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.

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