Part:BBa_K539461

promoter (lacI regulated)+RNA thermometer+vioD with RNA thermometer+tetR+double terminator

Please refer to the wiki for our overall concept:[http://2011.igem.org/Team:NCTU_Formosa/VP_design 2011 NCTU_Formosa]

We use the pathway which is mentioned by iGem 2010 team Slovenia, called the violacein biosynthesis pathway.

This pathway is a spontaneous cascade pathway which the initial compound, L-tryptophane is catalyzed by Vio A to indole-3-pyruvic acid imine (IPA imine) which then is converted into dimer by Vio B.
VioE then transforms the dimer to protodeoxyviolaceinic acid (PVA).
The resulting PVA could then further be converted into different products, such as deoxyviolacein which is catalyzed by VioC and the other one is protoviolaceinic acid by VioD.
The resulting protoviolaceinic acid can then again be converted into violacein by VioC.
We design our circuits this way to obtain the chain mechanism which at the end of the mechanism we have the accumulated protoviolaceinic acid. We then convert these PVA into violacein by using temperature controlled vioC expression.

Circuit B

Figure.1 Circuit B (Part:BBa_K539461 ,DH5α,PSB1C3): The expression of vioD

In circuit B (Part:BBa_K539461), the expression is promoted by Plac promoter (Part:BBa_R0010) but the expression is restricted by 37oC RBS (Part:BBa_K115002) which means, the ribosome will bind to the ribosome binding site only if the temperature reach 37 oC or higher, which the vioD (Part:BBa_K539413) is then translated.
The VioD then catalyze PVA into protoviolaceinic acid. The other translated sequence is tetR (Part:BBa_C0040), which repress the expression of circuit A which has Ptet (Part:BBa_R0040). This way, Ecoli will concentration on yielding vioD instead of wasting resources on unwanted product.

Sequence and Features

Assembly Compatibility:

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

  Illegal NgoMIV site found at 2119
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1194
  Illegal SapI.rc site found at 1269