Part:BBa_K801040

SV40NLS-PhyB-linker-Gal4DBD (part for GAL4 based light switchable promoter system)

Composite part of BBa_K801030 and BBa_K801038. One part of this fusion protein codes for the first of two components of a GAL4 based light-switchable promoter system. This GAL4DBD of this part is responsible for binding of GAL4 responsive elements like the upstream activating sequence (UAS) of pGAL1. The PhyB part is responsible for the direct recruitment of PIF3 (phytochrome interacting factor 3) and hence responsible for the indirect recruitment of the transcriptional activation domain GAL4AD because PIF3 is fused to GAL4AD (BBa_K801039).

It is known that there is already a endogenous nuclear localization signal in the N-terminus of GAL4DBD but to ensure nuclear translocation of the fusion protein, an additional SV40 nuclear localization is fused to the N-terminus in this part.

The second part, needed for a light-switchable promoter system, is BBa_K801039. There is also a complete expression casette battery for the two components of a GAL4 based yeast light-switchable promoter system, driven by TEF1 promoters (BBa_K801042).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 550
  Illegal BamHI site found at 632
  Illegal XhoI site found at 583
  Illegal XhoI site found at 602
  Illegal XhoI site found at 2737
  Illegal XhoI site found at 3071
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2122
  Illegal BsaI site found at 2528
  Illegal BsaI site found at 2990
  Illegal SapI site found at 799