Team:TUDelft/Parts/Main

Parts Overview

When constructing our BioBricks, we made use of the IDT synthesis offer. This way we were able to generate our required parts in a fast and efficient way. Additionally, we made use of PCR amplification with our constructed expression plasmids as template to construct several BioBricks. All of the parts contain a coding sequence, some of which extended with promoters or introns. Since our fusion protein BBa_K2643000 consists of the dxCas9, linker and Tn5 coding sequences this part is registered as composite part. The basic parts comprising the our fusion protein composite part are documented separately and also exist as separate BioBricks. This is advantageous for those wishing to combine one of our basic parts with a different expression level or compose a different composite part altogether. These basic parts were generated by using PCR on the composite part and restriction ligation cloning. We created a part collection comprised of 6 basic parts encoding for different EPO coding sequences, 2 basic parts encoding for the required components for targeted sequencing and 1 composite part: our fusion protein. All BioBricks are submitted with elaborate characterisation so the user may choose the best protein for their project accordingly. Our favourite basic part is BBa_K2643001 and our favourite composite part is BBa_K2643000.

Table 1. BioBrick Parts submitted by TU Delft iGEM 2018 team. All BioBricks were constructed by the team members involved in wetlab: Susan Bouwmeester, Kavish Kohabir, Venda Mangkusaputra, Timmy Paez, Lisbeth Schmidtchen

Fave Part	Name	Type	Description	Designer
♥	BBa_K2643000	Composite	dxCas9-linker-Tn5 fusion protein (HIS Tag)	TU Delft 2018
♥	BBa_K2643001	Basic	dxCas9 (HIS Tag)	TU Delft 2018
♥	BBa_K2643002	Basic	Tn5 Transposase (HIS Tagged)	TU Delft 2018
	BBa_K2643003	Basic	Glycine Helical peptide Linker (GHL	TU Delft 2018
♥	BBa_K2643004	Basic	Coding region (CDS) of human erythropoietin (EPO) hormone	TU Delft 2018
	BBa_K2643005	Basic	Human EPO gene with artificial intron (615 bp)	TU Delft 2018
	BBa_K2643006	Basic	Human EPO gene with artificial intron (135 bp)	TU Delft 2018
	BBa_K2643007	Basic	Human EPO gene with 2 artificial introns (615 + 135 bp)	TU Delft 2018
	BBa_K2643008	Basic	Human EPO gene with mutation 1 (5 bp)	TU Delft 2018
	BBa_K2643009	Basic	Human EPO gene with mutation 2 (12 bp)	TU Delft 2018
	BBa_K2643010	Basic	gRNA expression cassette for CRISPR-targeting lacZ	TU Delft 2018
	BBa_K2643011	Basic	Mosaic Ends-flanked Kanamycin cassette and RFP	TU Delft 2018
	BBa_K2643012	Basic	T7p expressing sgRNA targeting the EPO coding sequence	TU Delft 2018