Part:BBa_K1783003

Unstable RFP under Hok/Sok Regulation

Produces unstable RFP under a constitutive promoter. This construct carries the hok/sok toxin/antitoxin (TA) plasmid maintenance system and was used to measure the ability of this alternative plasmid maintenance system to keep plasmids within the cell without the use of antibiotics. LVA-tagged unstable RFP has a half life of 1 hour allowing real-time data observations.

Usage and Biology

We used this to test the ability of Hok/Sok to maintain plasmids over a long period of time. Hok/Sok is used naturally in the cell to maintain the R1 plasmid. RFP is a natural fluorescence protein found from coral.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 587
  Illegal NheI site found at 610
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1199
  Illegal AgeI site found at 1311
  
• 1000
  COMPATIBLE WITH RFC[1000]

Hok/Sok is directly adjacent to the RFP construct. This may be modified in future work by adding an insert in between Hok/Sok and RFP in order to decouple possible promoter interference.