Part:BBa_K4237022
PvanR_THS
Improve van induced promoter. RNA appratus is added downstream PvanCC
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 90
Illegal NheI site found at 237 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 66
Illegal BamHI site found at 104
Illegal BamHI site found at 213
Illegal BamHI site found at 251 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
This a an improvement part of Part:BBa_K3317006
THS strategy
The THS regulator encodes a structural component followed by a ribosome binding site (RBS) that is used to drive translation of the GOI. The structural region is designed to form a strong hairpin loop that when transcribed hinders the ability for ribosomes to bind the RBS, and thus inhibits translation. Translation is activated by expression of a complementary small RNA (sRNA) trigger that hybridizes to a short unstructured region of the THS, causing a breakdown in its secondary structure. This conformational change allows ribosomes to bind the RBS and translation of the GOI to proceed. THSs were selected because they offer strong repression of translation, can be designed computationally, and large libraries of designs exist with minimal crosstalk when used together
Figure. 7 Characterization of PVan_CC and PVan_CC_THS.
We tested the response of the food additive vanillic acid response promoter PvanCC and found that its activation ploidy was low compared to the original literature, and to further increase its activation ploidy, we used the literature-reported, RNA-coordinated THS strategy to increase the activation ploidy from approximately 20-fold to 34-fold.
Figure. 7 Characterization of PVan_CC and PVan_CC_THS.
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