Part:BBa_K2933169

Tac promoter+RBS a+Linker g+GST+Linker e+MUS-2

This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+MUS-2),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
  COMPATIBLE WITH RFC[21]
• 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

This composite part is made up with four basic parts(Tac promoter，RBS a , Linker g , GST+Linker e+MUS-2).It encodes a protein which is MUS-2 fused with GST tag. The fusion protein is about 54.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine the two parts with the cutting site of Prescission Protease. The fusion protein can be cut off at the cutting site by Prescission Protease. It is convenient for us to purify our target protein.

Molecular cloning

We used the vector pGEX-6p-1 to construct our expression plasmid.

Figure 1. Left: The PCR result of MUS-2. Right: The verification results by enzyme digestion.

After verification, it was determined that the construction is successful.