Difference between revisions of "Part:BBa K2992027"

 
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<partinfo>BBa_K2992027 short</partinfo>
 
<partinfo>BBa_K2992027 short</partinfo>
  
<i>botR</i> integration module for <i>C. sporogenes</i> with the lactose inducible system <i>bgaRL</i>
 
  
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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 P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] whose gene is a cognate member 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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===Characterisation===
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Data incoming
  
 
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<partinfo>BBa_K2992027 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2992027 SequenceAndFeatures</partinfo>
  
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===References===
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Cañadas et al., 2019 RiboCas – update
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Dupuy  and Matamouros, 2006 update
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Hartman and Melville 2011 - update
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Minton et al., 2016 Road map – update
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Raffestin et al 2009 update
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 11:35, 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 ofC. 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


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