Part:BBa_K1453201

[ssDsbA]->[RFP]->[Lgt]->[TAL-adapter]

Overview

This part is the fundamental unit of our project. It consists of four subunits: ssDsbA, RFP, Lgt, and TAL-adapter. ssDsbA is the signal recognition particle (SRP) - dependent signaling sequence of DsbA. ssDsbA-tagged proteins are exported to the periplasm through the SRP pathway. Lgt is an inner membrane protein of E.coli with seven transmembrane segments and has been successfully over expressed in E. coli without causing harm to cells. So with the help of ssDsbA and Lgt, we can make our fusion protein anchored in the membrane. RFP is the reporter, it can be detected by microscope. And the most important part is the TAL-adapter. We put two restriction endonuclease sites at the beginning and end of the adapter. Our part and 2012 Freiburg TALE parts, after digestion by BsmBI and ligation by T4 ligase, become a functional DNA binding protein which mostly exists in the membrane.

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1703
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1703
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1703
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1703
  Illegal AgeI site found at 612
  Illegal AgeI site found at 724
  Illegal AgeI site found at 1537
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1158