Part:BBa_K2923019

MS2-MS2-Theophylline inactive 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 protein and RNA. To make the interaction between BH3 and aptazyme iGEM Strasbourg created an aptazyme bordered by MS2 and PP7 stem-loops.

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

This sequence corresponds to inactive aptazyme: the ribozyme module is mutated and the aptazyme lost self-cleavage capability. iGEM Strasbourg 2019 used it as a negative control for the part: BBa_K2923018

Figure 2: Schematic graphic of inactive aptazyme. Inactive aptazymes (lost self-cleavage capability) active BH3 system independently of ligand presence

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]