Part:BBa_K1820019

CP44_RBS_mCherry(Lr)_Terminator

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Usage and Biology

This is a composite part designed as a report protein sequence for use in Lactococcus lactis. The promoter is a high-range promoter 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 codon optimized for Lactobacillus reuteri and biobricked according to standard 25, and a popular double-stop terminator. 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 cell and two other similar constructs (see Figure 1).

Figure 1. Fluorescence levels from three constructs using synthetic promoters from Jensen and Hammer (1998) in E. coli excited at 530/25 with emissions read at 590/35. cp8 from part BBa_K1820017, cp11 from part BBa_K1820018, and cp44 from part BBa_K1820019

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]

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.