Part:BBa_K2483000

IAA producing enzymes fused to RNA-binding proteins under control of Pveg2

This is a fusion protein of IaaM (Indoleacetic Acid tryptophan monooxygenase (IaaM) BBa_K515000) catalysing the oxidative carboxylation of L-tryptophan to indole-3-acetamide and MCP a RNA-binding protein from the virus MS2. Additionally, we've included the pVeg2 promoter and rbs (BBa_K5151010) so this part is essentially the part K515100 of Team Imperial College 2011 without the IaaH and with MCP fused to IaaM.

We used this part (in conjunction with BBa_K2483001) to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by puttign them in close proximity to each other (metabolic channelling). This was done with a dCas9 scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of sgRNA and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are located behind each other (BBa_K2483005 and BBa_K2483006).

There is another part with IaaH fused to PP7, another RNA-binding protein called BBa_K2483001. For the complete part with sgRNA, dCas9 and the IAA-enzymes, see BBa_K2483004.

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 547
  Illegal BamHI site found at 1492
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 254
  Illegal NgoMIV site found at 3198
  
• 1000
  COMPATIBLE WITH RFC[1000]