Part:BBa_K1499004

Cellulose binding domains with streptavidin domain generator

This part encodes a protein with two cellulose-binding domains on either end joined by a streptavidin domain.

Usage and Biology

The idea behind this part is that each domain binds to a single cellulose fiber, thus providing a way to cross-link and strengthen cellulose polymers. In addition, the streptavidin domain allows for the modular addition of biotinylated proteins through protein-protein interactions.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 703
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 304
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 651
  

Characterization

Verification of Part

The part was sequence verified before submission to the registry with two reads using VF2 and VR.

Figure 2. The forward read using VF2.

Figure 3. The reverse read using VR.

Results

We plan to assay the functionality of this part using a GFP construct inside a cell with the AviTag/OmpA surface complex. We will purify and apply our CBD/Streptavidin fusion protein to a cellulosic surface, followed by the above transformed cells, and attempt to wash off the cells using an isotonic solution. If the level of fluorescence does not change, we can assume our system has been successful.