Team:Imperial College/Basic Part





Basic Part

pSoxS Mutant 3

pSoxS is one half of the pSoxR/pSoxS bidirectional promoter. Transcription downstream of pSoxS is activated in response to oxidation of the SoxR transcription factor, either directly by redox-cycling drugs or by oxidative stress. It is therefore inducible by various redox-cycling drugs, toxins, antibiotics, heavy metals, hydrogen peroxide and nitric oxide, providing various applications in the development of environmental and therapeutic devices. By coupling oxidation of redox-cycling species to an electrode, the 2018 Imperial College London iGEM team (PixCell) used pSoxS to build electrogenetic devices in which electrical inputs modulated gene expression. This pSoxS promoter represents an improvement on Part:BBa_K387001. Our primary contribution is enhanced modularity, allowing its incorporation into a library of promoters and corresponding transcription factors; we also have evidence of new functionality (transcriptional repression) in combination with one of the transcription factors in this library. The promoter was taken from E. coli, with a 2bp deletion introduced between the -35 and -10 site in order to convert the induction of the promoter from transcriptional activation to transcriptional repression. The promoter was also engineered to be unidirectional by knocking out activity of the pSoxR portion. It was designed with an upstream terminator to remove the need for these to be added as parts in large assemblies. It also contains a downstream ribozyme to reduce context-dependency. It forms part of the PixCell library of electrogenetic and redox-sensing parts. This part is compatible for BioBrick, BASIC and Golden Gate assembly.
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