Difference between revisions of "Part:BBa K2992048"

 
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<partinfo>BBa_K2992048 short</partinfo>
 
<partinfo>BBa_K2992048 short</partinfo>
  
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FAST reporter construct with P<i>j23106</i> and <i>thl</i> RBS
 
  
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===Usage and Biology===
 
===Usage and Biology===
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This parts entry represents a FAST reporter construct under the regulatory control of P<i>j23106</i> from <i>E. coli</i>. The construct comprises the promoter P<i>j23106</i> [https://parts.igem.org/Part:BBa_J23106 BBa_J23106] coupled with the RBS from the <i>C. acetobutylicum</i> thiolase gene [https://parts.igem.org/Part:BBa_K2715009 BBa_K2715009], driving the expression of the Fluorescence-Activating and Absorption-Shifting Tag protein (FAST) reporter gene [http//parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcriptional terminator occurs through the activity of the strong clostridial terminator T<i>Fad</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus <i>Clostridium</i>. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.  <br><br>
 
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===Characterisation===
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Data incoming.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2992048 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2992048 SequenceAndFeatures</partinfo>
  
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===References===
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Heap modular
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Street et al 2019. Update
  
 
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Revision as of 13:26, 1 October 2019


FAST reporter construct with Pj23106 and thl RBS


Usage and Biology

This parts entry represents a FAST reporter construct under the regulatory control of Pj23106 from E. coli. The construct comprises the promoter Pj23106 BBa_J23106 coupled with the RBS from the C. acetobutylicum thiolase gene BBa_K2715009, driving the expression of the Fluorescence-Activating and Absorption-Shifting Tag protein (FAST) reporter gene [http//parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcriptional terminator occurs through the activity of the strong clostridial terminator TFad BBa_K2284012. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus Clostridium. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.

Characterisation

Data incoming.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 231


References

Heap modular Street et al 2019. Update