Part:BBa_K2992044

FAST reporter construct with PntnH 5-UTR+RBS.

Usage and Biology

This parts entry represents a FAST reporter construct under the regulatory control of PntnH for the non-toxic non-haemagglutinin ntnH gene of C. botulinum. The construct comprises the strong clostridial promoter PntnH BBa_K2992001 and its associated 5’-UTR containing the RBS BBa_K2992015 driving the expression of the fluorescent reporter gene FAST 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
  COMPATIBLE WITH RFC[12]
• 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 330
  

References

Heap modular Street et al 2019. Update