Part:BBa_K1075043

pLac-RBS32-mazE-(Ec)ssrA(DAS+4)-TT

Structure

This part consists of pLac2-RBS32-mazE-(Escherichia Coli)ssrA(DAS+4)-TT

Biology

MazE is part of the biological toxin-antitoxin system MazEF. This system is derived from the bacterium Escherichia coli, where it serves as a stress inducable killswitch. MazE is the antitoxin of this part and normally inhibits the activity of MazF. Absence of MazE leads to cell death, lest it be expressed in high levels during a certain timeframe [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC532418/]. The Escherischia Coli ssrA tag is part of the ClpXP system of E. Coli. In this system, it serves as a degradation tag that is recognized either by sspB or the ClpXP protease, both resulting in the degradation of the tagged protein. The DAS+4 mutation decreases the sensitivity of the ClpXP protease for the ssrA tag, thus increasing the dependency of mediation via sspB for degradation [http://www.ncbi.nlm.nih.gov/pubmed/16762842].

Application

This part can be used to gain a Lactose and IPTG inducable construct of mazE-(ec)ssrA(DAS+4). We used this construct as part of our proof of concept.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 524
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 274
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]