Part:BBa_K4885007:Design

1

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 214
  

Design Notes

In the butyrate synthesis pathway of C. tyrobutyricum L319, crotonase (crt) converts 3-hydroxybutyryl-CoA to crotonyl-CoA which is subsequently converted to butyryl-CoA by butyryl-CoA dehydrogenase coded by bcd. These two enzymes are rate-limiting enzymes. We used this part and BBa_K4885008 to express fusion protein of NGTag and crotonase (crt) and fusion protein of NGCatcher and butyryl-CoA dehydrogenase (bcd) in C. tyrobutyricum, which allows the assembly of crotonase and butyryl-CoA dehydrogenase via the NGTag/NGCatcher peptide interaction. In this way, we were able to enhance the butyrate synthesis pathway in the strain.

Source

NGCatcher sequence is from BBa_K4119013 bcd sequence is from BBa_K4119027.