Part:BBa_K5382130

CL7-linker-sfGFP_Green fluorescent protein and CL7 complex

The CL7 is a protein tag engineered from colicin CE7. In E.coli, CE7 is a bacteriocin that belongs to the colicin DNase family and has endonuclease activity. Through engineering, CL7 removed DNA binding and catalytic activity, but retained its high affinity binding ability to the corresponding inhibitory protein Im7. This property makes CL7 a useful tool for protein purification and molecular biology applications.
The linker here is used to connect CL7 and sfGFP short peptide sequence.
The sfGFP, namely the super folding green fluorescent protein (Superfolder GFP), is a genetically engineered green fluorescent protein (GFP) variations.
In the experimental design, we took advantage of the high affinity between CL7 and Im7 and incubated CL7-Linker-sfGFP to bind to Inak-linker-Im7(another composite part we registered), thus anchoring the integrated system to the membrane surface. Finally, the Inak-linker-Im7-CL7-linker-sfGFP system was demonstrated on the membrane surface, where the function of sfGFP protein was tested by double fluorescence verification (see the experimental design section for details) to see whether the system was successfully displayed on the membrane surface. It should be noted that the system we designed uses the high affinity of CL7 and Im7, so some components can be flexibly replaced according to needs, and are suitable for different use scenarios according to different needs. For example, according to different cells to replace the ice nucleated protein, according to different targeting targets to replace sfGFP with different single-chain antibodies, nano antibodies and so on.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 158
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 158
  Illegal NheI site found at 1247
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 158
  Illegal BglII site found at 106
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 158
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 158
  Illegal AgeI site found at 56
  Illegal AgeI site found at 70
  
• 1000
  COMPATIBLE WITH RFC[1000]