Part:BBa_K1616002

VVD linked to YN155 (YFP Nter split) with double terminator T7

Vivid (VVD) is the smallest known Light–oxygen–voltage (LOV) domain protein and photo-inducible dimer. Isolated from Neurospora crassa, VVD forms a homodimer in response to a blue-light stimulus. Then, a split protein is a protein whose sequence has been divided into two (or more) different parts. The yellow-fluorescent (YFP) protein will only express fluorescence when its two parts will be reunited. The part is coding for the homodimer VVD links by an integration of specific sequence to the N terminal of the YFP split. The downstream part of this composite is double T7 terminator (BBa_B0015).

So, this part works with BBa_K1616001. In absence of blue-light, the conformation of the VVD photoreceptor will prevent the formation of the complete fluorescent protein while in presence of the light signal the YFP protein will be reconstituted leading to the fast expression of a yellow fluorescence in our bacteria.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The sequence of VVD had 2 illegal sites PstI; that have been removed. Also, the N terminal of YFP had 1 illegal site PstI

Source

This part have been created thank to gblock, our team have assembled the sequence of photoreceptor VVD (without illegal site), a linker(1) and then the N terminal of YFP split(1). The downstream part of this composite part is double T7 terminator (BBa_B0015) and the upstream is the RBS (BBa_J61100).

References

(1) Tom Kerppola, Ph. D, investigator at the Howard Hughes Medical Institute as well as Professor in the University of Michigan

Hu CD, Chinenov Y, Kerppola TK. Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. Mol Cell. 2002;9(4):789–98.