Regulatory

Part:BBa_K2194002

Designed by: Catherine Dunaway, Anna Guseva   Group: iGEM17_Rice   (2017-10-23)
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PgntK Negative Feedback Membrane Stress Promoter

The Pgntk promoter is a native Escherichia coli negative feedback membrane stress promoter. BioBrick BBa_K2194002 contains the 200 bp upstream of the gntK gene in Escherichia coli, which has been previously experimentally shown to incorporate the full Pgntk promoter region. [1]


Biology and Usage

PgntK is native promoter of the gluconate metabolism operon in E. coli. It helps mitigate stress imposed by the overexpression of membrane proteins by providing negative feedback regulation of the membrane protein genes. It has been demonstrated that PgntK can optimize expression levels of AcrB drug efflux pump to maximize the growth rate and minimize cellular toxicity [1].


[1] Boyarskiy, Sergey et al. “Transcriptional Feedback Regulation of Efflux Protein Expression for Increased Tolerance to and Production of N-Butanol.” Metabolic Engineering 33 (2016): 130–137. Web. 28 Oct. 2017.


Pgntk.png

Figure 1: PgntK (BBa_K2194002) Promoter similar strength to Pcon (BBa_J23108) Figure 1: we compared the reporter protein expression of E. coli MG1655 transformed with a PgntK promoter+mCherry construct versus a Pcon promoter+mCherry construct. The data indicates that PgntK promoter has a comparable transcription initiation rate to Pcon promoter when no membrane stress is present. 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 site found at 14
    Illegal BsaI.rc site found at 216


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