Part:BBa_K2923020

MS2-MS2-Guanine actif aptazyme-PP7-PP7

Our bacterial three-hybrid (BH3) construct (inspired by Berry and Hochschild, 2018) is composed of two fusion proteins with MS2 and PP7 coat proteins and RNA. To make the interaction between BH3 and guanine aptazyme iGEM Strasbourg created an aptazyme bordered by MS2 and PP7 stem-loops. This aptazyme is specific for the guanine or guanosine ligand. It is composed of 3 different parts: a ribozyme module, a communication module, and an aptamer module. This aptazyme is a switche-on type of aptazyme. Its catalytic activity (self-cleavage) requires the presence of ligand.

Figure 1: Plasmid overview of MS2-MS2-Aptazyme-PP7-PP7

This aptazyme is a switche-on type of aptazyme. Its catalytic activity (self-cleavage) requires the presence of ligand.

Figure 2: Schematic graphic of Guanine aptazyme switch-on based bacterial three-hybrid system

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 37
  Illegal PstI site found at 25
  Illegal PstI site found at 64
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 25
  Illegal PstI site found at 64
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 37
  Illegal PstI site found at 25
  Illegal PstI site found at 64
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 37
  Illegal PstI site found at 25
  Illegal PstI site found at 64
  
• 1000
  COMPATIBLE WITH RFC[1000]