Part:BBa_K1075026
AraC-pBad-RBS34-mCherry-(Ec)ssrA(DAS+4)-TT
The part contains the AraC-pBad promoter system, a ribosomal binding site, the red fluorescent protein mCherry fused to the E. coli(ec)ssrA (DAS+4) tag and a double terminator.
biology
The (ec)ssrA tag is a short peptide sequence that can be fused to the C-terminus of proteins which should be degraded. The adaptor protein SspB binds to it and delivers the substrates to the protease ClpXP complex in E. coli. The mutation (DAS+4) in (ec)ssrA prevents the degradation of the ssrA-tagged protein without the presence of sspB and therefore makes the system more specific and controllable. [http://www.ncbi.nlm.nih.gov/pubmed/16762842]
mCherry is a red fluorescent protein with the excitation maximum at 587 nm and the Emission maximum at 610 nm. [http://www.ncbi.nlm.nih.gov/pubmed/15558047]
application
As we want to control protein degradation by controlling the function of ecSspB, we tagged the red fluorescent protein mCherry with ecssrA(DAS+4) to measure the degradation rate.
The part was designed for proof of principle. Application as a bacterial fotographic film might be possible as well.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2726
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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