Part:BBa_K3590041

Enzyme MlrB derived from Sphyngopyxis sp. fused with a dockerin derived from Bacteroides cellulosolv

MlrB is the second enzyme in the microcystin degradation pathway. The purpose of the MlrB-DocBc is to degrade the product of the reaction catalyzed by MlrA into even shorter linear peptides and thus contributing to microcystin degradation. This enzyme should fulfill its task while being displayed on ScafD.

At the 5' end of MlrB-DocBc there is the Pveg promoter followed by the RBS R1. The CDS encodes several functional modules - SacB signal sequence for extracellular transport, His-tag for Western blot detection followed by the MlrB module, the function of which is described above. MlrB is connected by a short linker to a dockerin of Bacteroides cellulosolvens. This dockerin interacts exclusively with a cohesin domain originating from the same species ensuring the attachment of this fusion protein to the specific position on ScafD. At the 3' end of the CDS, there is a STOP codon.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1769
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1325