Part:BBa_K1486054:Design

CheY/CheZ + split rLuc

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1497
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1205
  Illegal PstI site found at 1497
  
• 21
  INCOMPATIBLE WITH RFC[21]

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

  Illegal PstI site found at 1497
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1497
  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 961
  

Design Notes

When designing the part, we took care of adding a start codon at the beginning of the C terminus part of the split luciferase and deleting the stop codon at the end of the same sequence.

The two fusion proteins for the experiment mentionned in the main page are on the same plasmid. They are separated by a short random sequence followed by a second RBS.

Source

Both parts of the split were obtained from plasmid pYNZC from Waldor laboratory [http://waldorlab.bwh.harvard.edu/] used in this paper [http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0043175].

CheY sequence was obtained from part BBa_K569017 [[1]] from UT Dallas 2011 team.

CheZ sequence was obtained from part BBa_K629003 [[2]] from SYSU China 2011 team.

References

S.K. Hatzios, S. Ringgaard, B. M. Davis, M. K. Waldor (2012, August 15). Studies of Dynamic Protein-Protein Interactions in Bacteria Using Renilla Luciferase Complementation Are Undermined by Nonspecific Enzyme Inhibition Plos One.