Part:BBa_K3477004

Gene coding for his-tagged BphA2 enzyme including BBa_0034 RBS, Bba_J23106 promoter and Bba_0010 ter

This part constitutes the translational unit of Biphenyl dioxygenase subunit alpha 2 (BphA2). 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, BphA2, originates from Pseudomonas sp. KKS102 and takes part in the first step of the Biphenyl degradation pathway, in combination with BphA1. The Biphenyl degradation pathway degrades biphenyl molecules to benzoate and 2-oxopent-4-enolate. 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 758
  Illegal EcoRI site found at 1
  Illegal XbaI site found at 56
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal NheI site found at 27
  Illegal NheI site found at 50
  Illegal SpeI site found at 759
  Illegal PstI site found at 773
  Illegal NotI site found at 7
  Illegal NotI site found at 766
  
• 21
  INCOMPATIBLE WITH RFC[21]

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

  Illegal suffix found in sequence at 759
  Illegal EcoRI site found at 1
  Illegal XbaI site found at 56
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 1
  Illegal XbaI site found at 56
  Illegal SpeI site found at 759
  Illegal PstI site found at 773
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 456