Part:BBa_K4685000

SpyCatcher fused to gene Als3 ( Rubber-binding protein)

This composite part, includes the rubber-binding protein fused to the SpyCatcher protein. The SpyCatcher-RB complex is capable of interacting with SpyTag domain fused to the CsgA(BBa_K4685021) (main component of the Curli fiber) . The SpyTag/SpyCatcher system allows an irreversible conjugation of proteins: the peptide SpyTag reacts with the protein SpyCacther and forms an intramolecular isopeptide bond between the two proteins. We decided to leverage this key-lock mechanism by combining the SpyCatcher with the Rubber-binding protein and the SpyTag with the csgA subunit. Upon reaction between the two parts, the Curli fibers would be decorated with the Rubber-binding domain

Figure 1: Schematic representation of the SpyCatcher/SpyTag system applied to our project. The SpyTag protein, located on the surface of Curli fiber, will interact with the SpyCatcher protein that is bound to the Rubber-binding protein.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 1462
  Illegal PstI site found at 361
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 1462
  Illegal PstI site found at 361
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1427
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 1462
  Illegal PstI site found at 361
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 1462
  Illegal PstI site found at 361
  Illegal NgoMIV site found at 1161
  Illegal AgeI site found at 1294
  Illegal AgeI site found at 1579
  
• 1000
  COMPATIBLE WITH RFC[1000]