Part:BBa_K1616022

pDawn - Endolysin/Holin

The plasmids pDawn for light-activated gene expression. Basically, the YF1/FixJ drives gene expression from the pFixK2 promoter in a blue-light repressed manner. When we insert the λ phage repressor cI and the λ promoter pR in pDawn, this will invert the signal polarity and lead to the gene expression.

The histidine kinase YF1 employs a light oxygen voltage blue light photosensor domain. In the absence of blue light, YF1 phosphorylate the regulator FixJ which induces gene expression from the FixK2 promoter. The opposite happens with pDawn.

Toxins will be produced after light stimulation and induce lysis.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1405
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2755
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2689
  Illegal NgoMIV site found at 2787
  Illegal NgoMIV site found at 3670
  Illegal NgoMIV site found at 3742
  Illegal NgoMIV site found at 3832
  Illegal NgoMIV site found at 3850
  Illegal NgoMIV site found at 4344
  Illegal AgeI site found at 306
  Illegal AgeI site found at 376
  Illegal AgeI site found at 3384
  Illegal AgeI site found at 4512
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4401
  Illegal BsaI.rc site found at 3283
  

Design Notes

We obtained the plasmids from the Centre for Biological Signalling Studies and the University of Freiburg. As soon as we received them, we observed that the multiple cloning site as well as the λ promoter pR were located upstream the YF1/FixY system.

Source

This part is composed of Holin/Endolysin toxin BBa_K1616005 and pDawn system BBa_K1616019