T7

Part:BBa_K5250003

Designed by: Ekaterina Tocheva   Group: iGEM24_UZurich   (2024-09-28)


PT7

The T7-like promoter os a promoter derived from the Pseudomonas phage Pf-10. It recruits the Pseudomonas phage Pf-10 RNA polymerase (BBa_K5250002) which after binding triggers expression.


This serves as a non-toxic alternative to the T7 Escherichia coli T7 RNA polymerase-T7 promoter system in Pseudomonas species. 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


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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