Part:BBa_K339010:Design
ibpAB
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Source
Designed from literature sequences by Kraft et al., 2007.
Derivation of the promoter
This promoter is found naturally in E. coli. This promoter is comprised of two naturally occuring promoters.
This image shows the plasmid schematic from paper by Kraft et al.
In order to derive the promoter that is submitted, ibpAB (native) was blasted in order to locate the location in the sequence provided in the paper.
Clustal W alignment with native promoter region in front of ibpAB operon(P2_ibpAB)and the sequence provided in the paper (P1_proposed)
Similarly, the FxsA promoter (native) was also aligned with the entire sequence in the paper.
These two alignments allowed us to derive the important DNA parts. There were parts eliminated. For example an internal PvoII site was eliminated as well at a T7 ribosome binding site was removed to use standard ribosome binding site. (B0034)
References
Kraft, M., Knupfer, U., Wenderoth, R., Pietschmann, P., Hock, B., & Horn, U. (2007). An online monitoring system based on a synthetic sigma32-dependent tandem promoter for visualization of insoluble proteins in the cytoplasm of escherichia coli. Applied Microbiology and Biotechnology, 75(2), 397-406.