Composite

Part:BBa_K1163102

Designed by: Emiel van der Kouwe   Group: iGEM13_Evry   (2013-09-22)
Revision as of 23:20, 4 October 2013 by Eccho (Talk | contribs)

pAceB-sfGFP-Term
iGEM Evry 2013

Description:
This part is composed of the following elements:

  • AceB promoter region
  • superfolder GFP
  • Terminator

The AceB gene encodes the malate synthase A enzyme, which is involved in carbon source management. Its promoter region has been extracted from genomic Escherichia coli DNA. After genomic research, we found that the gene is under the regulation of fur, thus meaning it is sensitive to iron concentration variations.

FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In the presence of iron, FUR binds iron and dimerizes. This modification of conformation binds to DNA on a Fur Binding Site and inhibits the mRNA transcription of the downstream gene.
This 300 bp long promoter sequence contains a RBS and a FUR binding site, although it has not been clearly possible to identify them.

Characterization
To caracterize our promoter sequence, we cloned a superfolder GFP downstream. It has been shown to downregulate the expression of sfGFP or any gene down-stream in the presence of iron in the range: 10-7 to 10-4 mol.L-1 Why would you use this part? This part allows the characterization of the AceB promoter region thanks to the sfGFP downstream. Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 277
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 319


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Parameters
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