Part:BBa_K2992025
botR integration module for C. sporogenes with native promoter, 5-UTR+RBS
Usage and Biology
This parts entry represents an integration module for the expression of botR from the pyrE locus of the C. sporogenes genome. This module comprises a strong clostridial terminator sequence Tfad to prevent polar transcription from pyrD BBa_K2992013 and the botR gene of C. botulinum BBa_K2992002, under the regulatory control of its native promoter PbotR BBa_K2992012 and its associated 5’-UTR which contains the RBS BBa_K2992014. An additional strong clostridial terminator was included to prevent polar transcription of pyrE and any downstream genes on the chromosome of C. sporogenesBBa_K2284012. In our project we use the transcriptional regulator of neurotoxin production from C. botulinum, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST BBa_K2992000 through interaction with its own promoter sequence PbotrR BBa_k299012 and PntnH BBa_K2992001 whose genes are cognate members of the BotR regulon. Doing so allows us to use our surrogate host strain C. sporogenes as a model system for predicting botulinum neurotoxin production following food manufacture, through the detection of our chosen reporters.
Characterisation
[[File:FAST data.png]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
Cañadas et al., 2019 RiboCas – update Dupuy and Matamouros, 2006 update Minton et al., 2016 Road map – update Raffestin et al 2009 update
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