Intermediate

Part:BBa_K1075041

Designed by: Marc Schulte   Group: iGEM13_Bonn   (2013-09-23)

RBS32-MazE-(Ec)ssrA(DAS+4)

Structure

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

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 linked with promotors and different terminators to generate either inducible or constant expression of mazE-ecssrA(DAS+4). Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 316
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 66
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None