Parts

Introduction

Our team added 9 new basic parts and 6 new composite parts to the iGEM registry. As mentioned, our project is divided into three main designs: Compete, Attack, and Protect. Thus we have three main groups of parts, in accordance with our designs.

The parts that appear in bold were characterized.

Best Basic Part

Our best part is paraE (BBa_K4633006) it was characterized by measuring fluorescence in B. subtilis and in E. Coli, and by imaging E. coli.

Compete

The goal of this design is to allow Lactobacillus crispatus to compete with UPEC on its adhesion sites, thus preventing UPEC from establishing in the microbiota and re-starting the infection.

Detailed information regarding the new parts we introduced into the registry is presented in the table below:

Part name	Part number	Part type	Basic / Composite	Length	Picture
FimHL - Mannose-Binding Domain of FimH	BBa_K4633000	Coding	Basic	489
AmyA - signal peptide for Lactobucilli	BBa_K4633001	Protein domain	Basic	114
AmyL - signal peptide for Lactobucilli	BBa_K4633002	Protein domain	Basic	114
Pveg-AmyE-Based Platform for FimHL and GFP Co-expression in B. subtilis	BBa_K4633100	Composite	Composite	1432
Pveg-AmyE-Based Improved Platform for FimHL and GFP Co-expression in B. subtilis	BBa_K4633101	Composite	Composite	1471

Attack

The primary objective of this design is to enhance our effectiveness against UPEC by attacking it. It is based on the Colicin D toxin, which we added as a part, and existing parts from past iGEM groups.

A summary of the parts for this system can be found in the following table:

Part name	Part number	Part type	Basic / Composite	Length	Picture
Colicin D - Bacteriocin from E. coli	BBa_K4633008	Coding	Basic	2091

Protect

The primary objective of this design was to prioritize user safety. As detailed in our protect page, it became evident that there were no relevant parts in the iGEM registry suitable for our specific requirements. We sought an inducible system that could be triggered by arabinose, since unlike IPTG it is safe for human consumption, and could induce expression in a Gram-positive bacterium.

The table below provides a concise overview of the parts associated with this design:

Part name	Part number	Part type	Basic / Composite	Length	Picture
araR - Bacillus subtilis repressor protein	BBa_K4633003	Coding	Basic	1089
paraR - promoter for the araR gene from Bacillus subtilis 168	BBa_K4633004	Regulatory	Basic	55
RBS paraR - the RBS from the promoter for the araR gene from Bacillus subtilis 168	BBa_K4633005	RBS	Basic	12
paraE - improved promoter for the araE gene from Bacillus subtilis 168	BBa_K4633006	Regulatory	Basic	113
RBS paraE - the RBS from the promoter for the araE gene from Bacillus subtilis 168	BBa_K4633007	RBS	Basic	14
paraR-mCherry	BBa_K4633102	Composite	Composite	781
paraE-mCherry	BBa_K4633103	Composite	Composite	841
Bacillus subtilis L-arabinose induced system	BBa_K4633104	Composite	Composite	2077
Bacillus subtilis L-arabinose induced killswitch	BBa_K4633105	Composite	Composite	1702