Part:BBa_K3933018

Standardized version of our sensor to be adapted for sequestering riboswitches

This part was developed for our implemented Human Practice, so that other teams could be able to adapt our system to the molecule they want to detect. This construction contains a Riboswitch Insertion Zone (RIZ), which contains restriction enzymes sites. Thanks to these sites, this construction can be digested and used for Golden Gate Assembly or Restriction Enzyme assembly.

This part is composed of two units.

The first one is the sensing unit, composed of a constitutive promoter (BBa_J23106), followed by the RIZ (BBa_K3933010). The riboswitch needs to be inserted in the RIZ sequence for the part to be properly used. The riboswitch will work with the RBS which is placed just after (BBa_B0032) to regulate the expression of the Tet R protein (BBa_C0040). The last part of this unit is the double terminator (BBa_B0015). The second one is the signalling unit, composed of a Tet Operator (which reacts to Tet R expression) (BBa_R0040), a RBS (BBa_B0032) and a double terminator (BBa_B0015) framing a Lactate Dehydrogenase (BBa_K3102006).

To have more details on the functioning of this part can be found on the part initially used for our system : BBa_K3933007

Unfortunately, due to lack of time, this part could not be tested in the lab.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 142
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 46
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1553
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1283