Part:BBa_K5377840

TPP self-induced production system Ⅱ

In order to self-induced production of TPP(thiamine pyrophosphate) without the need for human intervention, our team designed this part. This composite part integrates the thiM TPP riboswitch 'ON system' and a TPP synthesis operon. It is mainly used to detect subtle fluctuations in intracellular TPP concentrations and intelligently regulate the TPP synthesis pathways through thiM TPP riboswitch. By adding the lacI gene after the riboswitch sequence and inserting the lacO gene before the operon, we transformed the riboswitch from an 'OFF system' that is unfavorable for production into the 'ON system' that we need.
When the concentration of the TPP is low, the thiM TPP riboswitch remains in its ON state, allowing the downstream lacI gene to produce a repressor protein that binds to the lacO operator, inhibiting the expression of the small molecule synthesis operon. As the ligand concentration rises and reaches a specific threshold, the riboswitch shifts to the OFF state, stopping the production of the repressor protein. This enables normal expression of the operon, leading to significant synthesis of the small molecule TPP.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 3747
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3747
  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3747
  Illegal BglII site found at 4184
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 3747
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 3747
  Illegal AgeI site found at 2742
  Illegal AgeI site found at 4074
  Illegal AgeI site found at 4305
  
• 1000
  COMPATIBLE WITH RFC[1000]