Part:BBa_K606036

pT7 SpoVG T7 SpoVG GFP T7terminator.

T7 RNA polymerase auto-amplifier system (T7 autoloop) for B. Subtilis and E. coli

Usage and Biology

This construct, the so-called T7 autoloop, uses positive feedback to create signal amplification.

The T7 RNA polymerase has a very specific promoter (pT7, 25 nucleotides) that can be recognize by no other polymerases. The system is really orthogonal as long as there is no polymerase leakeage from the transcription in the plasmid. Synthetic biology plasmids (here pSB1C3) use 4 standards terminators before the promoter to limit such leakage. You might need to add some others if you are working on another plasmid.

The stochastic leakeage of this system has been characterized (see the experiments page). Once the system is activated, the cell stop dividing and glows very high.

You can use this part directly to detect a tiny amount of T7 RNA polymerase in a cell (in the case of a cell infection for instance).

The RBS used here are for B. subtilis but the system works fine in E. coli as well (as shown in the experiments).

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 3598
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3374