Regulatory

Part:BBa_K3259001

Designed by: Tsuruta Soh   Group: iGEM19_TokyoTech   (2019-10-15)
Revision as of 02:14, 22 October 2019 by Crane (Talk | contribs)

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pCold-Rhl hybrid promoter

This promoter has a RhlR / PAI2(C4-HSL) complex binding site located at the upstream of the cold-sensitive promoter pCold (BBa_K1949000).

　pCold is used because it can produce the target protein at low temperature efficiently. There is publication stated that 90% of nascent proteins are proteins expressed by pCold at low temperature condition. The expression level of pCold decreases at around 37 degrees, but some leakage occurs.

However, suppression effect at 37 degrees are further strengthened by the suppressive effect of RhlO. RhlO is derived from BBa_R0071 and is induced from the binding of RhlR / PAI2. The project is also expected to be able to control gene expression by two different methods, namely inducers and environmental factors. pCold is sensitive to low temperature because the structure of 5'UTR located at the downstream of cspA promoter is unstable when is around 37 degrees and is stabilized around 18 degrees. Therefore, we decided to place RhlR / PAI2 at the upstream of pCold.


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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Parameters
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