Part:BBa_K2835006

His-tagged laccase with start codon (ATG), mutated to increase the specificity for SMX

This BioBrick codes for a mutated and codon optimized version of a laccase from Trametes versicolor, namely 1GYC (PDB ID). It contains mutated residues at points 162, 332 and 337, from Phenylalanine to Isoleucine. The aim of the mutations was to increase the enzyme's specificity and activity for sulfamethoxazole. It has been codon optimized for Pichia pastoris.

This BioBrick is an alternative part to BBa_K2835004, where the only difference between these parts is the present or absent of a start codon (ATG). This Biobrick is a basic part containing a start codon (ATG). This means that this part can be directly ligated (without addition of a start codon through PCR) into any standard iGEM vector and can, after assembly with a promoter, be expressed in a host of choice. The alternative BioBrick BBa_K2835003 is a protein domain not including a start codon, which makes it easy to fuse it to for example a secretion signal or to ligate it into a vector contaning genes which can be fused.

For Usage and Biology as well as characterization, see BBa_K2835004.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 411
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]