Part:BBa_K4885006

Pthl-adhE2-bcd-crt

This part is responsible for the expression of bcd and crt enzymes derived from Clostridium tyrobutyricum (C. tyrobutyricum) L319 and adhE2 gene derived from Clostridium tyrobutyricum with Pthl promotor. It consists of Pthl sequence (BBa K3443002), strong ribosomal binding site (RBS) for adhE2 sequence (BBa_K4408001), adhE2 sequence(BBa_K1462060), bcd sequence(BBa_K4119027)， crt sequence (BBa_K4119028), and the Cpa fdx terminator sequence (BBa_K3585002). adhE2 gene codes for the alcohol/aldehyde bifunctional dehydrogenase. Its role is to convert acyl-CoA to aldehyde then to alcohol in two reductive steps using NADH as cofactor. To introduce adhE2 in C. tyrobutyricum, we can construct a butanol synthesis pathway based on the native butyrate synthesis pathway. Crt gene codes for crotonase which converts 3-hydroxybutyryl-CoA to crotonyl-CoA in the butyrate synthesis pathway in C. tyrobutyricum. bcd gene codes for butyryl-CoA dehydrogenase which converts crotonyl-CoA to butyryl-CoA in the butyrate synthesis pathway in C. tyrobutyricum. The crotonase and butyryl-CoA dehydrogenase are rate-limiting enzymes in the butyrate and butanol synthesis pathway.

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 3367
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4546
  Illegal SapI.rc site found at 3496