Regulatory

Part:BBa_K4271010

Designed by: Ethan Ho   Group: iGEM22_Wego_Taipei   (2022-10-05)


rpoD sigma factor


ropD gene (BBa_K4271010), comprises N-terminal 69 amino acid for polyP binding (Yang et al., 2010), E. coli RpoD (1-555 a.a), C-terminal 4.2 region for UreA promoter recognition (BBa_K4271011)(Beier et al., 1998).

The rpoD sigma factor is a factor under the sigma-70 family and is a common sequence in E. coli. Activated by polyphosphate bondage, the sigma factor will be expressed and assist the RNA polymerase to bind to downstream ureA promoter (BBa_K4271011), thus expressing the mCherry (BBa_J18932) fluorescent protein for detection to determine engineering success. The usage of the rpoD sigma factor can be seen in BBa_K4271013 and BBa_K4271014.

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References

Yang, Z. X., Zhou, Y. N., Yang, Y., & Jin, D. J. (2010, June 11). Polyphosphate binds to the principal sigma factor of RNA polymerase during starvation response in Helicobacter pylori. Molecular Microbiology, 77(3), 618–627. https://doi.org/10.1111/j.1365-2958.2010.07233.x

Beier, D., Spohn, G., Rappuoli, R., & Scarlato, V. (1998, October). Functional analysis of theHelicobacter pyloriprincipal sigma subunit of RNA polymerase reveals that the spacer region is important for efficient transcription. Molecular Microbiology, 30(1), 121–134. https://doi.org/10.1046/j.1365-2958.1998.01043.x


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 693
    Illegal BglII site found at 741
    Illegal BamHI site found at 1448
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1519


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