Part:BBa_K2923018

MS2-MS2-Theophylline active 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 aptazyme iGEM Strasbourg created an aptazyme bordered by MS2 and PP7 stem-loops. Aptazyme is composed of 3 different parts, a twister ribozyme module, a communication module, and an aptamer module.

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

The described anti-theophylline aptazyme is an OFF-switch. Its catalytic activity (self-cleavage) requires the absence of ligand.

Figure 2: Schematic graphic of Theophylline aptazyme switch-off 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]