Composite

Part:BBa_K3147006

Designed by: Thomas Bessede   Group: iGEM19_Montpellier   (2019-10-14)
Revision as of 06:25, 14 October 2019 by Registry (Talk | contribs)

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mutant TEV (PE10) protease with TEV-cleavable maltose binding protein

The Montpellier 2019 team made this construction in order to be able to compare the basal activity of the TEV protease (BBa_ K1319004) with the mutant TEV protease called TEVPE10, we were able to find the mutant TEV sequences in this article [3]. This construction was used as a control within the Karma project presented by the iGEM Montpellier 2019 team. It produces a mutant TEV fused to an MBP with, in between a modified TEV cutting site to increase the mutant's activity. MBP increases the solubility of the fusion protein [1] preventing the aggregation of the protein of interest ; the sequence of the produced MBP does not have a signal peptide which keeps the protein in the cytosol. The TEV cutting site allows to separate MBP from TEV once the protein is produced, indeed the TEV protease is capable of self-cleavage [2] to remove MBP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 77
    Illegal SapI.rc site found at 1868


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