Parts

Big things have small beginnings.

~ Prometheus

This page gives an overview of all the parts that we used in our project. They have been divided into three categories: the composite parts that represent our fusion proteins for the genetic engineering method, the parts that encode the proteins needed for the post-insertion method and the basic parts that were used to build up the forementioned parts.

Composite Parts: Fusion proteins

We created a library of parts (BBa_ K5403012 - BBa_K5403020) that encode fusion proteins that are designed to functionalize the membrane of the bacterial membrane vesicles (BMVs) of M. smegmatis and E. coli with a protein, see Table 1. These fusion proteins all consist of an N-terminal domain that is known to exist in the membrane of the BMVs, a flexible linker (BBa_K5403002) and a C-terminal GFP (BBa_K5403997). GFP was chosen for initial characterization, but each part contains an NheI-restriction site that allows you to replace the GFP with a gene for any protein of your interest. Click on the Part ID to see more information on that specific part.

*Composite parts: Fusion proteins for the genetic engineering method*
Part ID	Description	Designers
BBa_K5403012	Fusion protein for OmpAN21-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403013	Fusion protein for OmpA-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403014	Fusion protein for OmpAtb-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403015	Fusion protein for membrane anchoring of GFP based on OmpAtb and OmpA	iGEM24_TU-Eindhoven
BBa_K5403016	Fusion protein for porinMspA-signal-sequence-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403017	Fusion protein for truncated porinMspA-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403018	Fusion protein for porinMspA-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403019	Fusion protein for RbsC-based membrane anchoring of GFP	iGEM24_TU-Eindhoven
BBa_K5403020	Fusion protein for FtsQ-based membrane anchoring of GFP	iGEM24_TU-Eindhoven

Composite Parts: Parts for post-insertion method

Two parts were created for the post-insertion method for the functionalization of the BMVs. These parts are shown in Table 2. Click on the Part ID to see more information on that specific part.

*Parts for the post-insertion method.*
Name	Type	Description	Designers
BBa_K5403005	Improved	Spycatcher with unique Cysteine at N-terminus	iGEM24_TU-Eindhoven
BBa_K5403021	Composite	GFP with N-terminal Spytag and C-terminal hexahistidine tag	iGEM24_TU-Eindhoven

Basic Parts

For the assembly of the composite parts and the parts for the post-insertion method, a range of basic parts was used, as shown in Table 3. Click on the Part ID to see more information on that specific part.

*Basic parts.*
Part ID	Type	Description	Designers
BBa_K5403002	Coding	Flexible GSGSGSGSAS linker with NheI restriction site	iGEM24_TU-Eindhoven
BBa_K5403003	Coding	N-terminal signal sequence of OmpA	iGEM24_TU-Eindhoven
BBa_K5403004	Coding	Outer membrane protein A (OmpA)	iGEM24_TU-Eindhoven
BBa_K5403006	Coding	N-terminal signal sequence of OmpAtb	iGEM24_TU-Eindhoven
BBa_K5403007	Coding	N-terminal transmembrane domain of PorinMspA of M. smegmatis	iGEM24_TU-Eindhoven
BBa_K5403008	Coding	Porin MspA of M. smegmatis missing 13 C-terminal amino acids	iGEM24_TU-Eindhoven
BBa_K5403009	Coding	Membrane channel porin MspA of M. smegmatis	iGEM24_TU-Eindhoven
BBa_K5403010	Coding	Protein RbsC of M. smegmatis missing 35 C-terminal amino acids	iGEM24_TU-Eindhoven
BBa_K5403011	Coding	Protein FtsQ of M. smegmatis	iGEM24_TU-Eindhoven
BBa_K5403997	Coding	GFP as a C-terminal domain for M. smegmatis fusion proteins	iGEM24_TU-Eindhoven
BBa_K5403998	Tag	Spytag	iGEM24_TU-Eindhoven
BBa_K5403999	Tag	Hexahistidine-tag (His-tag) with C-terminal STOP-codon	iGEM24_TU-Eindhoven