4xMREa 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 (K5317007) for cell-based metal detection.

To increase the efficiency of the activated MTF-1-responsive promoter, we constructed a synthetic promoter with multiple MREa sites. Searle and colleagues described in 1985 that at least two MREa sites are necessary for the zinc-induced expression of the downstream gene, here herpes simplex virus thymidine kinase. They also showed that the positioning of the MREs in the promoter sequence had little effect on the promoter efficiency but was increased with more MREa sites inserted.

Cloning

Theoretical Part Design

The promoter sequence contains four MREa sites with respective core consensus sites for binding of heavy metal-activated MTF-1, their positions, and in-between DNA sequence based on our MREwt promoter, registry entry K5317003.

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]

Characterization

The promoter was analyzed by composing a gene cassette where its placed upstream of the reporter gene EGFP to assess the promoter's functionality and metal-dependent efficiency based on the fluorescent signal. Please visit the K5317009 registry entry to view the results.

References