Part:BBa_K3447007

xynD, which produces xylanase

T7 polymerase stage 2 with pMag.

Usage and Biology

In our project, we divided T7 RNA polymerase into two parts, respectively called T7_NT (BBa_K3447008) and T7_CT (BBa_K3447007) and attached on the nMag and pMag. Under blue light irradiation, these two parts undergo specific blue light dependent heterodimerization and assemble into complete T7 RNA polymerase, which acts on PT7 promoter (BBa_K3447005) and activates downstream gene expression. And the process is reversible, that is, when blue light is stopped, the dimerized T7 RNA polymerase dedimers and loses its function.

Figure 1. Main principles of the molecular mechanism of T7 promoter

In our project, we use this part to form a complete T7 RNA polymerase by dimerizing it with T7_NT under blue light. (链接104+f09+104+f10+001+b17)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 216
  Illegal PstI site found at 226
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 216
  Illegal PstI site found at 226
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 216
  Illegal PstI site found at 226
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 216
  Illegal PstI site found at 226
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design

Source

We found this sequence data in the following reference.

• Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.
  
  

References

1. Armin Baumschlager, Stephanie K. Aoki, and Mustafa Khammash, Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for Precise Spatiotemporal Gene Expression Control, ACS Synth. Biol. 2017, 6, 2157-2167.