Part:BBa_K2992027
botR integration module for C. sporogenes with the lactose inducible system
Usage and Biology
This parts entry represents an integration module for the expression of botR at the pyrE locus of the C. sporogenes genome. This module comprises the botR gene of C. botulinum BBa_K2992002 under the regulatory control of the BgaRL lactose inducible system from C. perfringens wherein the divergent PbgaRBBa_K2992020 and PbgaL BBa_K2992023 promoter sequences, coupled with their native 5’-UTR and RBS sequences BBa_K2992022[ https://parts.igem.org/Part:BBa_K2992024 BBa_K2992024] respectively control the transcription of bgaR and botR. Transcription for bgaR is terminated by its native terminator sequence BBa_K2992021 whilst botR uses Tfdx derived from C. pasteurianum BBa_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 PntnH BBa_K2992001 whose gene is a cognate member 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
Data incoming
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 573
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 573
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 573
Illegal BglII site found at 633 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 573
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 573
- 1000COMPATIBLE WITH RFC[1000]
References
Cañadas et al., 2019 RiboCas – update Dupuy and Matamouros, 2006 update Hartman and Melville 2011 - update Minton et al., 2016 Road map – update Raffestin et al 2009 update
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