Part:BBa_K3477005

Gene coding for his-tagged PETase enzyme including BBa_0034 RBS, Bba_J23106 promoter and Bba_0010 te

This part constitutes the translational unit of polyethylene therephtalate hydrolase (PETase). Upstream of the coding region there is the biobrick prefix, the BBa_J23106 constitutive promoter (from the Anderson family) and the BBa_B0034 ribosome binding site (RBS). The gene sequence comes from Uniprot.org and is codon optimized for Escherichia coli. The sequence is followed by a 6x his-tag sequence. Downstream of the coding region there is the BBa_B0010 terminator and the biobrick suffix.

The enzyme coded for, PETase, originates from ideonella sakaiensis and degrades PET to MHET. The included his-tag allows for purification of the expressed protein using a his-tag purification kit.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal suffix found in sequence at 962
  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 341
  Illegal XbaI site found at 56
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 341
  Illegal NheI site found at 27
  Illegal NheI site found at 50
  Illegal NheI site found at 273
  Illegal SpeI site found at 963
  Illegal PstI site found at 977
  Illegal NotI site found at 7
  Illegal NotI site found at 970
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 341
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal suffix found in sequence at 963
  Illegal EcoRI site found at 1
  Illegal EcoRI site found at 341
  Illegal XbaI site found at 56
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 1
  Illegal EcoRI site found at 341
  Illegal XbaI site found at 56
  Illegal SpeI site found at 963
  Illegal PstI site found at 977
  
• 1000
  COMPATIBLE WITH RFC[1000]