Part:BBa_K2052016

FimH site directed mutated with RPMrel and ButCoat

........................... Enter a long description of the part so that users of your part know what it is, what it does, and how to use it in their projects.

This part is composed of a protein coding section (fimH+ RPMrel), a double terminator, arabinose induced promoter, RBS and another protein coding sequence (ButCoaT).

FimH protein is a subunit of a structure called fimH, which naturally occurs in some strains of e.coli. FimH adhesin is found at the end of each pilus, allowing bacteria to bind to sugar mannose. (Zeynep bitirince eklenicek)

RPMrel is a 9 amino acid a colon tumor specific binding heptapeptide. It was used for controlling preferential binding to poorly-differentiated colon carcinoma cells.

It includes a double terminator (BBa_B0015) consisting of BBa_B0010 and BBa_B0012. It works in the forward direction with a forward efficiency of 0.984[CC] and 0.97[JK].

BBa_K206000 is an arabinose induced e.coli promoter. We decided to work with Arabinose Induced Promoter in the middle of our construct to understand if FimH works without putting arabinose and if ButCoaT works with arabinose in the medium.

BBa_B0034 is a ribosome binding site with the efficiency of 1. It is responsible for the recruitment of the ribosomes during the initiation of protein translation.

Butyrate is a four carbon, short chain fatty acid that inhibits cancer. It induces apoptosis and differentiation, inhibits proliferation of tumorous cells in colon flora. It is produced by the bacterial fermantation of carbohydrates in colon. Butyrate is formed by many pathways which one of them begins with Acetyl-CoA. Acetyl-CoA is readily present in the cells so we wanted to find an enzyme that directly converts it to butyrate, which is ButCoaT. ButCoaT converts acetyl CoA to butyrate by the reaction Butanoyl-CoA + Acetate <=> Butanoic acid + Acetyl-CoA. And BBa_K2052018 is the sequence which codes for ButCoaT

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Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1173
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1113
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]