Team:OLS Canmore Canada/Parts

PARTS

Objective

Our objective is to insert this DNA construct into both E. coli and Bacillus subtilis to secrete proteins, which will fluoresce red and selectively bio-tag to PET plastic. We based our construct design on Tianjin’s 2016 PET-ase construct for the biodegradation of PET plastic. We modified this construct with the help of our mentor Lisa Oberding from Fredsense for this specific project. We need to pay huge attribution to our mentor Lisa who designed our construct and did many rounds of troubleshooting so we could finally get these parts synthesized. We chose Bacillus subtilis because of its natural ability to produce hydrophobins, and because when compared to E.coli, it is much better at naturally secreting proteins. The four constructs we designed for lab work all consist of two transcription terminators, a Bacillus-specific promoter, a ribosome binding site, which starts the formation of protein, and a LipA, which is a secretion tag that will help send the protein outside the cell. The parts we submitted to the registry are optimized for use in E.coli and are in the standard pSB1C3 backbone. Any teams wishing to get the sequences for Bacillus specific plasmid vectors are welcome to reach out to us for the sequence!

BBa_K2650001 and BBa_K2650003 (click for full size)

BBa_K2650001: mCherry PET-ase

BBa_K2650003: BsIA mCHERRY

BBa_K2650002: PET-ase

BBa_K2650002 and BBa_K2650004 (click for full size)

BBa_K2650004: BslA

Parts Table <groupparts>iGEM18 OLS_Canmore_Canada</groupparts>