Part:BBa_K1820018
CP11_RBS_mCherry(Lr)_Terminator
Usage and Biology
- This has only been tested in E. coli thus far.
This is a composite part designed as a report protein sequence for use in Lactococcus lactis. The promoter is a mid-range promoter, part BBa_K1033221 from the 2013 Uppsala team, from Peter Ruhdal Jensen and Karin Hammer's library of synthetic promoters for Lactococcus lactis followed by a popular ribosome binding site (Elowitz 1999), an mCherry fluorescent protein, part BBa_K1033250 from the 2013 Uppsala team, codon optimized for Lactobacillus reuteri and biobricked according to standard 25, and a popular double-stop terminator. We further characterized this promoter with the addition of a fluorescent protein. Although it was designed for L. lactis, it has displayed function in Escherichia coli as well. There are indications that the promoter is functional in a large number of prokaryotic organisms. As such, it is likely that this construct will be functional in a variety of prokaryotic organisms.
We created and tested this construct in Escherichia coli in the pSB1C3 plasmid. It was tested for fluorescence relative to non-transformed E. coli cells (see Figure 1).
This part improves upon CP11 (https://parts.igem.org/Part:BBa_K1033221) by the addition of a RBS, fluorescent protein, and a terminator, as well as further characterization of the promoter.
Figure 1. Fluorescence levels from BBa_K1820018 in E. coli excited at 530/25 with emissions read at 590/35
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
Jensen, P. R., Hammer, K. (1998). The Sequence of Spacers between the Consensus Sequences Modulates the Strength of Prokaryotic Promoters. Appl Environ Microbiol. 1998 Jan; 64(1): 82–87. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC124675/
mCherry Fluorescent Protein. Clonetek, Takara Bio. http://www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Proteins_by_Name/mCherry_Fluorescent_Protein. Accessed 17 Sept 2015.
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