Team:Linkoping Sweden/Basic Part

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Basic parts

Basic parts

Down below is our basic part collection. All basic parts were designed to be able to assemble with BBa_K3182001 and our expression system, BBa_K3182200. To be able to assemble our basic parts with our cellulose binding domain (CBD_cipA) we added a BamHI site inside the linker region of our construct, see Figure 1. The BamHI site was chosen because of it being a part of the RFC21, and coding for one glycine and one serine which is the end sequence of the thrombin cleavage site. The thrombin cleavage site was added into the linker region to be able to separate our fusion proteins. We had no trouble assembling our fusion proteins using the pink-white screening method (BBa_K3182100).

Figure 1: The construct is composed of a carbohydrate binding domain (CBD_cipA) with a GS-linker containing a thrombin (Th. CS) and BamHI site on the C-terminal (3'-end). The BamHI-site on the end of the linker allows exchange of fusion protein. The figure on the top is a picture of the construct in Benchling illustrating the CBD, GS-linker followed by a Th. CS and BamHI, ended by an agent which can be switched for any protein. The picture below is a graphical representation.

Basic parts

Table 2. Basic parts used in the wet lab part of our project. All basic parts used was designed to fit for assembly with BBa_K3182001.

Biobrick ID	Short name	Function	Size	Designed by
BBa_K3182001	CBDcipA	Carbohydrate binding domain with a C-terminal linker	516 bp	Oliver Hild Walett, Johan Larsson
BBa_K3182003	PlyF307-SQ8C	Acinetobacter genus lysin	444 bp	Oliver Hild Walett
BBa_K3182004	CHAP(K)	Staphylococcus genus lysin	489 bp	Leo Juhlin
BBa_K3182005	LL-37	Antibacterial peptide	117 bp	Oliver Hild Walett
BBa_K3182006	Magainin 2	Antimicrobial peptide	75 bp	Oliver Hild Walett
BBa_K3182007	Pln1	Antibacterial peptide	117 bp	Oliver Hild Walett
BBa_K3182009	Parseqα	Synthetic antimicrobial peptide	60 bp	Johan Larsson, Jonatan Baggman
BBa_K3182010	Parseqβ	Synthetic antimicrobial peptide	60 bp	Johan Larsson, Jonatan Baggman
BBa_K3182013	mNeonGreen	Fluorescent protein	771 bp	Oliver Hild Walett, iGEM Linköping 2018

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