Part:BBa_K4591012

P43-DepoPETase-MHETase-T500

Usage and Biology

This circuit contains the coding sequences of P₄₃, DepoPETase, and MHETase. The expression of enzyme genes related to this route is controlled by the frequently expressed promoter P43, which degrades PET into TPA and achieves the preliminary degradation of PET.

Basic components of the circuit

P₄₃：P₄₃ is a constitutive promoter that constitutively expresses the P₄₃ protein in B.subtilis.

DepoPETase：DepoPETase is the best mutant obtained by researchers using a novel high-throughput screening method based on fluorescence detection for directed evolution of IsPETase.

MHETase：MHETase is discovered from Ideonella sakaiensis 201-F6, it is a enzyme which can hydrolyze MHET, which is the main product of PETase when hydrolyzing PET.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 421
  Illegal PstI site found at 588
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 421
  Illegal PstI site found at 588
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 421
  Illegal XhoI site found at 505
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 421
  Illegal PstI site found at 588
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 421
  Illegal PstI site found at 588
  Illegal NgoMIV site found at 275
  Illegal NgoMIV site found at 2010
  Illegal AgeI site found at 1230
  Illegal AgeI site found at 1347
  
• 1000
  COMPATIBLE WITH RFC[1000]