Composite

Part:BBa_K3311005

Designed by: Shan Dong   Group: iGEM19_Worldshaper-Shanghai   (2019-06-16)
Revision as of 08:13, 21 October 2019 by SylviaSong (Talk | contribs)

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Improved HucR-pHucO-mCherry

It is an improvement on BBa_K3311004. We added a strong constitutive promoter (BBa_J23100) in front of pHucO, enhancing the transcription of mCherry. In addition, the original strong promoter before HucR is replaced by a weaker constitutive promoter (BBa_J23110) so as to weaken the repression on pHucO.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 759
    Illegal NheI site found at 782
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

This composite part was improved based on the suggestions from QHFZ-China 2019 iGEM team during an online meet up. The leader of their team told us that the pHucO we were using is too weak to let the gene pathway to produce enough mCherry. Plus, the promoter of HucR is too strong, causing a heaving repression on the transcription on mcherry. Furthermore, they provided two articles to support their suggestion[1,2]. According to their suggestion, we designed an improved plan: we added a strong constitutive promoter (BBa_J23100) in front of pHucO, enhancing the transcription of mcherry. In addition, the original strong promoter before HucR is replaced by a weaker constitutive promoter (BBa_J23110) so as to weaken the repression on pHucO.

Reference

1. Kemmer, C., Gitzinger, M., Daoud-El Baba, M., Djonov, V., Stelling, J., and Fussenegger, M. (2010) Self-sufficient control of urate homeostasis in mice by a synthetic circuit, Nature biotechnology 28, 355-360.

2. Wan, X., Volpetti, F., Petrova, E., French, C., Maerkl, S. J., and Wang, B. (2019) Cascaded amplifying circuits enable ultrasensitive cellular sensors for toxic metals, Nature chemical biology 15, 540-548.

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