Part:BBa_K5250018

Prha + DGC WspR R242A

In this transcriptional unit the wild type DGC of Pseudomonas aeruginosa is expressed after a rhamnose promoter (BBa_K914003).

Usage and Biology

In our project this is one of the biofilm overexpression modules we are testing. DGC, or diguanylate cyclase, produces the second messenger cyclic di-GMP which is involved in the biofilm production of bacteria. When DGC is overexpressed, this results in the induced formation of biofilm. The mutation WspR R242A is supposed to inhibit the negative feedback mechanism c-di-GMP has on the DGC activity. We want to increase the activity of DGC and the levels of c-di-GMP inside the cell and thereby boost biofilm production [1].

[1] De, N., Pirruccello, M., Krasteva, P. V., Bae, N., Raghavan, R. V., & Sondermann, H. (2008). Phosphorylation-Independent Regulation of the Diguanylate Cyclase WSPR. PLoS Biology, 6(3), e67. https://doi.org/10.1371/journal.pbio.0060067

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 894
  Illegal PstI site found at 336
  Illegal PstI site found at 561
  Illegal PstI site found at 633
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 894
  Illegal PstI site found at 336
  Illegal PstI site found at 561
  Illegal PstI site found at 633
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 894
  Illegal BamHI site found at 1215
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 894
  Illegal PstI site found at 336
  Illegal PstI site found at 561
  Illegal PstI site found at 633
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 894
  Illegal PstI site found at 336
  Illegal PstI site found at 561
  Illegal PstI site found at 633
  Illegal NgoMIV site found at 258
  Illegal NgoMIV site found at 1160
  Illegal NgoMIV site found at 1166
  
• 1000
  COMPATIBLE WITH RFC[1000]