Part:BBa_K2560262

mcrN - N-terminal domain of Malonyl-CoA Reductase from Chloroflexus aurantiacus

This part contains the 5' part of the mcr gene for Malony-CoA Reductase from C. aurantiacus, encoding the C-terminal domain. This enzyme is involved in the 3-hydroxypropionate cycle for CO2 fixation and converts malonyl-CoA into 3-hydroxy propionic acid (Strauss & Fuchs, 1993). In a two-step reaction malonyl-CoA is first reduced to 3-oxopropanoic acid and then further reduced to 3-hydroxypropionic acid. The C-terminal domain catalyzes the first reaction step to 3-oxopropanoic acid and the N-terminal domain catalyzes the second reaction step to 3-hydroxypropionic acid.

It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take this part and BBa_K2560261. If the gene for the whole enzyme shall be used take BBa_K2560260.

For this part, mcrN was codonoptimized for V. natriegens and then synthetisized and integrated into the vector BBa_K2560002 via BsmBI. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013).

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 885
  Illegal BamHI site found at 213
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1188
  Illegal AgeI site found at 1270
  
• 1000
  COMPATIBLE WITH RFC[1000]