Difference between revisions of "Part:BBa K3697009"

Revision as of 04:33, 25 October 2020

pVeg YFP Plasmid for B. subtilis

This part is an expression vector for Bacillus subtilis that produces codon-optimized YFP, for integration into Bacillus subtilis at the AmyE loci (BBa_K143001). YFP 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.

YFP has an excitation peak is 514 nm and an emission peak is 527 nm.

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

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

The YFP does not have a degradation tag.

Sequence and Features