Part:BBa_K2387055

Split mRFP controlled by inducible araC/pBAD promoter

This a split version of BBa_K2387054. The two halves of mRFP are fused to leucine zippers to induce the reassembly. Both parts are under the control of the araC/pBad promoter and each part is under the control of a stron RBS. The reassembly process is irreversible. This part can be used as a positive control for experiments using BiFC, as well as a source for the fragments of mRFP.

The mRFP has already been split for the iGEM Registry (BBa_I715022 & [BBa_I715023]), where the mRFP was split at the position 155. However, in these parts the mRFP is not split at the right site, and therefore does not produce fluorescence upon interaction. As observed by Guido et al. (2006), mRFP only is able to reassemble into a functional fluorescent protein when it is split in the position 168.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  Illegal AgeI site found at 2043
  Illegal AgeI site found at 2155
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961
  

References

Guido, Jach, et al. "An improved mRFP1 adds red to bimolecular fluorescence complementation." Nature Methods 13.8 (2006): 597-600.