Part:BBa_K2483000

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

This part is an upgrade to the part BBa_K515100 (https://parts.igem.org/Part:BBa_K515100) of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively.

We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (https://parts.igem.org/Part:BBa_K2483002) scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of a sgRNA and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are consecutively arranged (https://parts.igem.org/Part:BBa_K2483005 and https://parts.igem.org/Part:BBa_K2483006).

For the complete part with sgRNA, dCas9 and the IAA-enzymes, see https://parts.igem.org/Part: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]