Part:BBa_K1616019
pDawn system
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.
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 26
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1376
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1310
Illegal NgoMIV site found at 1408
Illegal NgoMIV site found at 2291
Illegal NgoMIV site found at 2363
Illegal NgoMIV site found at 2453
Illegal NgoMIV site found at 2471
Illegal NgoMIV site found at 2965
Illegal AgeI site found at 2005
Illegal AgeI site found at 3133 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3022
Illegal BsaI.rc site found at 1904
Design Notes
The original sequence of pDawn have some illegal sites that have been removed.
Penn iGEM 2015 characterized the pDawn system as a light-sensitive receiver in its light-mediated cell communication system. With mRFP cloned downstream of the light-responsive promoter, the system's light-mediated gene expression was confirmed. Furthermore, the system was activated using small amount of artificial light produced with an LED. These small amounts reflect the intensity of light output from luminescent sender cells. For additonal information, follow this link: http://2015.igem.org/Team:Penn/Receiver.
Source
The pDawn & pDusk system, have been optimized and constructed in the way that the toxins sequences we introduce in our construction are in fact the reverse sequences.
References
Ohlendorf, Robert, Roee R. Vidavski, Avigdor Eldar, Keith Moffat, and Andreas Möglich. "From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression." Journal of Molecular Biology 416.4 (2012): 534-42. Web.
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