Difference between revisions of "Part:BBa K3697010"

Revision as of 00:12, 24 October 2020

mCherry_BSU Plasmid

This part is an expression vector for Bacillus subtilis that produces codon-optimized mCherry_BSU, for integration into Bacillus subtilis at the AmyE loci (BBa_K143001). mCherry_BSU is under pVeg expression (BBa_K143012), the strongest constitutive promotor known in B. subtilis. Plasmid expresses kanamycin resistance in B. subtilis, and ampicillin resistance for cloning in E. coli.

mCherry_BSU has an excitation peak at 585 nm and a peak emission at 615 nm

This plasmid successfully integrates into B. subtilis and produces mCherry_BSU at levels that can be quantified using a fluorescent plate reader. Transformation was performed using xylose inducible competent B. subtilis, and plated on selective kanamycin media (20ug/mL). When cloning in E. coli, the E. coli may be red, as they will non-discriminately express the mCherry_BSU. E. coli transformed with this plasmid have been clearly red under natural light for the duration of their lifetime and does not appear to degrade. Transformed B. subtilis colonies may appear weakly red under natural light.

The RBS for expression of mCherry in this vector is strong.

The mCherry does not have a degredation tag.

Sequence and Features