Difference between revisions of "Part:BBa K2992027"
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===Usage and Biology===
 
===Usage and Biology===
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This parts entry represents an integration module for the expression of <i>botR</i> at the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises the <i>botR</i> gene of <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002] under the regulatory control of the BgaRL lactose inducible system from <i>C. perfringens</i> wherein the divergent P<i>bgaR</i> [https://parts.igem.org/Part:BBa_K2992020 BBa_K2992020] and P<i>bga</i>L [https://parts.igem.org/Part:BBa_K2992023 BBa_K2992023] promoter sequences, coupled with their native 5’-UTR and RBS sequences [https://parts.igem.org/Part:BBa_K2992022 BBa_K2992022]/[ https://parts.igem.org/Part:BBa_K2992024 BBa_K2992024] respectively control the transcription of <i>bgaR</i>  and <i>botR</i>. Transcription for <i>bgaR</i> is terminated by its native terminator sequence [https://parts.igem.org/Part:BBa_K2992021 BBa_K2992021] whilst <i>botR</i> uses T<i>fdx</i> derived from <i>C. pasteurianum</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. In our project we use the transcriptional regulator of neurotoxin production from <i>C. botulinum</i>, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000] through interaction with its own promoter sequence P<i>botrR</i> [https://parts.igem.org/Part:BBa_K2992012 BBa_k299012] and P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] whose genes are cognate members of the BotR regulon. Doing so allows us to use our surrogate host strain <i>C. sporogenes</i> as a model system for predicting botulinum neurotoxin production following food manufacture, through the detection of our chosen reporters. <br><br>
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This parts entry represents an integration module for the expression of <i>botR</i> at the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises the <i>botR</i> gene of <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002] under the regulatory control of the BgaRL lactose inducible system from <i>C. perfringens</i> wherein the divergent P<i>bgaR</i> [https://parts.igem.org/Part:BBa_K2992020 BBa_K2992020] and P<i>bga</i>L [https://parts.igem.org/Part:BBa_K2992023 BBa_K2992023] promoter sequences, coupled with their native 5’-UTR and RBS sequences [https://parts.igem.org/Part:BBa_K2992022 BBa_K2992022] and [ https://parts.igem.org/Part:BBa_K2992024 BBa_K2992024] respectively control the transcription of <i>bgaR</i>  and <i>botR</i>. Transcription for <i>bgaR</i> is terminated by its native terminator sequence [https://parts.igem.org/Part:BBa_K2992021 BBa_K2992021] whilst <i>botR</i> uses T<i>fdx</i> derived from <i>C. pasteurianum</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. In our project we use the transcriptional regulator of neurotoxin production from <i>C. botulinum</i>, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000] through interaction with its own promoter sequence P<i>botrR</i> [https://parts.igem.org/Part:BBa_K2992012 BBa_k299012] and P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] whose genes are cognate members of the BotR regulon. Doing so allows us to use our surrogate host strain <i>C. sporogenes</i> as a model system for predicting botulinum neurotoxin production following food manufacture, through the detection of our chosen reporters. <br><br>
  
 
===Characterisation===
 
===Characterisation===

Revision as of 11:42, 18 September 2019


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 PbgaR BBa_K2992020 and PbgaL BBa_K2992023 promoter sequences, coupled with their native 5’-UTR and RBS sequences BBa_K2992022 and [ 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 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

Data incoming

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 573
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 573
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 573
    Illegal BglII site found at 633
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 573
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 573
  • 1000
    COMPATIBLE 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