Composite

Part:BBa_K1163102

Designed by: Emiel van der Kouwe   Group: iGEM13_Evry   (2013-09-22)
Revision as of 23:13, 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.

This promoter sequence contains a RBS and a FUR binding site, although it is not clearly possible to identify them precisely. 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

FUR (Ferric Uptake Regulation) is a transcriptional repressor of genes involved in iron homeostasis. In presence of iron FUR will be linked to the iron. This modification of conformation will induce a dimerization of FUR. Then the dimeric FUR will bind to the DNA in a Fur Binding Site and inhibit the mRNA transcription.

To caracterize our constructions we use this Fur binding site merged with a super fold GFP to the power of inhibition of the Fur binding site.

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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Categories
Parameters
None