Part:BBa_K2933253

RBS b+Linker h+His+Linker a+Sumo+Linker b+GIM-2+T7 terminator

This part consists of RBS, protein coding sequence(His+Linker a+Sumo+Linker b+GIM-2) 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
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 298
  Illegal PstI site found at 1128
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 298
  Illegal NheI site found at 75
  Illegal NheI site found at 1158
  Illegal PstI site found at 1128
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 298
  Illegal BglII site found at 187
  Illegal BamHI site found at 386
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 298
  Illegal PstI site found at 1128
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 298
  Illegal PstI site found at 1128
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

This composite part is made up with eight basic parts, T7 Ribosome binding sites,the His-Sumo tag, three cutting sites of Prescission Protease, our target protein GIM-2 and T7 terminator. It encodes a protein which is GIM-2 fused with His-Sumo tag. The fusion protein is about 39.4 kD. The fusion protein can be cut off at the cutting sites by Prescission Protease. It is convenient for us to purify our target protein.

Experimental results

Molecular cloning

Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification