Protein_Domain

Part:BBa_K1075008

Designed by: Max Schelski   Group: iGEM13_Bonn   (2013-09-22)

E. coli sspB[Core]

This part contains the Core part of the split version of sspB.

Usage and Biology

EcsspB itself regulates the degradation of ssrA tagged proteins through the ClpXP protease in procaryotes. In engineered systems it is used to induce degradation of specifically ssrA tagged proteins.

This construct is part of the split system of E. Coli sspB (EcsspB) which was used in fusion with FKBP and FRB to induce the activity of sspB and therefore degradation of proteins with Rapamycins. EcsspB can be functionally divided in three parts: -the N terminal Core domain (113 AA) -the C terminal XB peptide (25 AA) -and a 'flexible linker' in between the first parts (28 AA) We used the same domain structure as is used within the Rapamycin inducable split system. While the Core domain is responsible for dimerization of sspB and binding of ssrA the XB peptide binds the protease ClpXP. The flexible linker is called flexible because it was found, that an increase or reduction in size or amino acid composition does not influence the function of sspB as much as it would in the other domains.

For our project we needed the sspB split system to engineer a light inducable sspB.

There is no stop codon included in this part, because regulatory parts were fused C-terminally to the XB peptide.


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
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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