Part:BBa_K801041

SV40NLS-PhyB-linker-LexA (part for a LexA based light-switchable promoter system)

Composite part of BBa_K801030, BBa_K801031 and BBa_K801036. One part of this fusion protein codes for the first of two components of a LexA based light-switchable promoter system. The LexA of this part is responsible for binding of LexA operator sequence upstream from a minimal promoter hence upstream from a TATA box, e. g. BBa_K165031, a part coding for the minimal promoter of CYC1 preceded by multiple consecutive binding sites for the LexA binding domain. 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).

In contrast to the GAL4 based promoter system (BBa_K801042, BBa_K801040), there is no endogenous nuclear localization signal in the binding domain, like in GAL4DBD, so to ensure nuclear translocation of the fusion protein, an additional SV40 nuclear localization is fused to the N-terminus of 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 LexA based yeast light-switchable promoter system, driven by TEF1 promoters (BBa_K801043).

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 BglII site found at 3390
  Illegal BamHI site found at 632
  Illegal XhoI site found at 583
  Illegal XhoI site found at 602
  Illegal XhoI site found at 2737
  
• 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 SapI site found at 799