Regulatory

Part:BBa_K5317005

Designed by: Jan Gelhoet   Group: iGEM24_Hannover   (2024-09-13)
Revision as of 09:44, 13 September 2024 by Annaseidler (Talk | contribs) (Usage and Biology)


4xMREd promoter

Usage and Biology

The MRE-sites containing promoter enables the metal-dependent expression of a downstream positioned reporter gene via the metal ion-dependent transcription factor MTF-1 for cell-based metal detection.

To achieve a cell-based heavy metal sensor, our group generated multiple synthetic MTF1 responsive promoter constructs and tested them in regard to their efficiency. Since Wang and colleagues (2004) suggested a high affinity of MTF-1 towards MREd we synthesized a promoter sequence containing four MREd elements at the positioning of the MREs of the MREwt promoter [1] to exclude possible disruption of the MTF-1 and MRE interaction.

Cloning

Theoretical Part Design

The sequence of the MREd site was obtained from Seguin and colleagues (1987), as well as the positioning and spacer sequences (Searle et al. 1985). The construct was synthesized.

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]


[edit]
Categories
Parameters
None