Regulatory
yenbox_J23

Part:BBa_K1381002

Designed by: Stephanie Herman, Gunta Celma, Megha Biradar, Christoffer Andersson and Martin Friberg   Group: iGEM14_Uppsala   (2014-10-02)
Revision as of 18:35, 17 October 2014 by StephyHe (Talk | contribs)

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yenbox_J23101

This part was constructed in the same way as BBa_K1381001 (yenbox_J23113). It consists of the promoter BBa_J23101 (J23101) from the Anderson promoter library fused together downstreams of a luxbox homolog, called the yenbox. The yenbox is a recognition region from Y. enterocoliticas quorum sensing system, the Yen system. The yenbox is recognised by an activator, BBa_K1381004 (YenR) and when interaction between these two occur, an induction of the promoters expression occur.

While designing this part, we tried to mimic the wildtype version. Hence, the wildtype version has an overlap of two bases between its promoter and the yenbox. Since the two last bases of the yenbox and the two first bases of J23101 did not match, we were faced with the choice of either changing two bases in the promoters sequence or in yenbox's sequence. In the article by Ching-Sung Tsai and Stephen C. Winanas[1] they discovered that the binding between the activator YenR and the recognition region of the yenbox is not dependent on the entire sequence of the yenbox. Depending on which part of the yenbox that is changed or replaced, YenR binds to the yenbox with different strengths. However, it still interacts with the yenbox and induces the strength of the promoter. Based on this fact, together with the knowledge that the Anderson promoters are very sequence dependent, we chose to change two bases in the sequence of the yenbox.

There has not been any characterisation experiments performed on this part. A construct for measurements has been assembled BBa_K1381010 (yenbox_J23101-B0032-GFP). However, yenbox_J23113 has been shown not to work (see characterisations of BBa_K1381001). It seemed like the activator YenR could not recognise nor interact with the yenbox due to the changes made in the last two bases of the yenbox.


References
[1] Ching-Sung Tsai and Stephen C. Winanas, The quorum-hindered transcription factor YenR of Yersinia enterocolitica inhibits pheromone production and promotes motility via a small non-coding RNA, 2011, Molecular Microbiology 80[2], 556-571


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 52
    Illegal NheI site found at 75
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None