Parts

Here, we present the parts we have created and, for most of them, validated during the course of this project. The main parts have been tested in multiple contexts (acellular, in liposome, in liposome in contact with cancer cells). While these were not designed to be a true collection, they offer a good basis to develop system biology approach using liposome.

To construct our parts, we used different approaches. Most gBlocks were designed on benchling and synthesized by IDT (which allowed us to obtain codon-optimized sequences for Escherichia coli), We got the other parts through PCR amplification, using instructions provided by another research team (you can find more details on the material page and in the wet lab results). The parts were then built in the desired vector by In-Fusion assembly.

Here is a summary table of all the parts iGEM Toulouse INSA-UPS 2023 designed this year. Key information is added such as length, the current status (works, failed, successful cloning, or in progress), and RFC compliance (parts with "*" cannot be associated with a RFC according to restriction site rules. Nonetheless, each individual element constituting these parts can be associated with at least one RFC).

If you want to know more about our parts or any inquiry, feel free to contact us at igem.toulouse@gmail.com.

• New parts:

Type	Purpose	Accession number	Nickname	Diagram	Significance in the project	Length (bp)	Status	RFC
Basic	Expression and purification of our transcriptional repressor DhdR for biosensing	BBa_K4768000	DhdR		***	973	Worked	12
Basic	Biosensing inducible system to express sfGFP in PURE system	BBa_K4768001	sfgfp		***	915	Worked	10, 12, 23, 25
Basic	Expression and purification of Nanobodies anti-HER2 for the anchoring to the liposome	BBa_K4768002	anti-Her2 nb		***	673	Worked	1000
Basic	Biosensing inducible system to express human thymidine phosphorylase in PURE system	BBa_K4768003	tymp		***	1647	Succesful cloning	10, 12, 23, 1000
Basic	Biosensing inducible system to express E. coli nucleoside phosphorylase (ppnP) in PURE system	BBa_K4768004	ppnP		***	466	Sequence validated	10, 12, 21, 23, 25, 1000
Basic	Expression of the T7 RNA polymerase Nterm conjugated to Pertuzumab with a soluble linker	BBa_K4768005	T7Nterm-SL-Pertuzumab		*	2057	Successful expression in PURE system	12, 21
Basic	Expression of the T7 RNA polymerase Cterm conjugated to Trastuzumab with a soluble linker	BBa_K4768006	Trastuzumab-SL-T7Cterm		*	4001	Cloning failed	12, 21
Basic	Expression of the T7 RNA polymerase Cterm conjugated to Trastuzumab with a transmembrane linker ZipA	BBa_K4768007	Trastuzumab-TM-T7Cterm		***	4103	Modeling	12, 21
Basic	Expression of the T7 RNA polymerase Nterm conjugated to Pertuzumab with a transmembrane linker ZipA	BBa_K4768008	T7Nterm-TM-Pertuzumab		***	2153	Modeling	*
Basic	Expression of the T7 RNA polymerase Nterm conjugated to rapamycin antibody (FRB) with a soluble linker	BBa_K4768009	T7Nterm-SL-FRB		*	982	Successful expression in PURE system	12
Basic	Expression of the T7 RNA polymerase Cterm conjugated to rapamycin antibody (FKBP) with a soluble linker	BBa_K47680010	FKBP-SL-T7Cterm		*	2596	Successful expression in PURE system	12, 21

• Improved parts:

Type	Purpose	Accession number	Diagram	Significance in the project	Length (bp)	Status	RFC
Basic	DhdR repressor binding site from Achromobacter denitrificans NBRC 15125	BBa_K4046100		***	75	Worked	10, 12, 21, 23, 25, 1000