Regulatory

Part:BBa_K5097000

Designed by: Kelly Acen, Marie Box, Alexia Darby, Jena Dookie, Blake Dieckman, Runya Chaora, Chris Dorce, Adrianna Dugan, Elber Lopez-Hernandez, kayla VanPelt, Makenna Ventuleth   Group: iGEM24_Oneonta   (2024-09-30)
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pH Response Element riboswitch

Usage and Biology

In 1990, Bingham et al. identified a locus within E.coli whose activity was tied to pH changes, which they coined Alx. It exhibited increased expression at pH 8.5 (1). This activity was further characterized by Nechooshtan et. al., who determined that the Alx promoter and riboswitch are responsible for increased Alx expression in response to high pH (2). The Alx promoter and riboswitch have been previously entered into the parts repository by Team NAXI_GRAS under part number BBa_K2348000.

Our team redesigned the pH-responsive element (PRE) from the Alex locus by isolating the riboswitch sequence to allow regulation of the riboswitch independent of the promoter. We utilized the PRE sequence published by (3) to delineate the riboswitch sequence. We did this so that an alternative promoter could be used with this part.


References

1-Bingham, R J, et al. “Alkaline Induction of a Novel Gene Locus, ALX, in Escherichia Coli.” Journal of Bacteriology, U.S. National Library of Medicine, Apr. 1990, www.ncbi.nlm.nih.gov/pmc/articles/PMC208722/.

2-Nechooshtan, Gal, et al. “A PH-Responsive Riboregulator.” Genes & Development, U.S. National Library of Medicine, 15 Nov. 2009, www.ncbi.nlm.nih.gov/pubmed/19933154.

3-Pham, H., Wong, A., Chua, N. et al. Engineering a riboswitch-based genetic platform for the self-directed evolution of acid-tolerant phenotypes. Nat Commun 8, 411 (2017). https://doi.org/10.1038/s41467-017-00511-w


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 118


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