Part:BBa_K5250002
T7-like RNAP
The T7-like RNAP is an RNA polymerase derived from the Pseudomonas phage Pf-10.
Usage and Biology
It is used as a non-toxic alternative to the T7 Escherichia coli RNA polymerase in Pseudomonas species. It binds to a the T7-like promoter of the same Pf-10 Pseudomonas phage. The T7 RNAP/pT7-complex from E. coli is widely used in synthetic biology and biotechnology for recombinant expression and is a valuable tool in microbiology. Therefore a similar construct in species outside of E. coli is highly beneficial. The sequence mentioned was chosen specifically to have low toxicity towards our target strain, high transcriptional activity and a low cross-recognition with the native to Pseudomonas RNAPs (1).
In the context of our project a T7-like RNAP has been used to build a positive feedback loop that further enhances the expression of our gene of interest. Furthermore, the T7-like RNAP links the biofilm overexpression module to the expression of our toxin system. It is used in multiple composite parts, both in our final construct and in various testing plasmids.
(1) Lammens, E.-M.; Feyaerts, N.; Kerremans, A.; Boon, M.; Lavigne, R. Assessing the Orthogonality of Phage-Encoded RNA Polymerases for Tailored Synthetic Biology Applications in Pseudomonas Species. Int. J. Mol. Sci. 2023, 24, 7175. https://doi.org/10.3390/ijms24087175
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 109
Illegal BsaI site found at 1173
Illegal BsaI site found at 2043
Illegal BsaI.rc site found at 61
Illegal BsaI.rc site found at 553
Illegal BsaI.rc site found at 1481
Illegal BsaI.rc site found at 2491
Illegal SapI site found at 2472
Illegal SapI.rc site found at 1667
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