Basic Parts Composite Parts in vitro characterization immobilization on our biofilm our kill switch

Due to Covid-19 we were not able to test any of our designed parts in the lab. Nevertheless, we registered our basic parts, so future teams are easily able to characterize them. For our Composite Parts we only uploaded one for which we can provide modeling data. We refrained from uploading the others to the registry as there exists no primary literature on them, but we still provide schematic images on them.

Basic Parts

Due to Covid-19 we were not able to test any of our designed parts in the lab. Nevertheless, we registered our basic parts, so future teams are easily able to characterize them. For our Composite Parts we refrained from uploading them to the registry as there exists no primary literature on them, but we still provide schematic images on them. All our basic parts are depicted in the table below. Because we were not able to conduct any lab work , we did not submit any parts for the special prizes Best Basic Part.

Part(BBa_)	Type	Description	Length in bp
BBa_K3429000	Promoter	P_grac promoter	170
BBa_K3429001	Coding Sequence	TasA matrix protein	783
BBa_K3429002	Coding Sequence	superfolder Green Fluorescent Protein (sfGFP)	712
BBa_K3429003	Coding Sequence	Protein Linker for fusion proteins in B. subtilis: ARGGGGSGGGGSGS	42
BBa_K3429004	Terminator	trpA-terminator	24
BBa_K3429005	Coding Sequence	Ribosomal Protein S2 (rpsB) from B. subtilis	721
BBa_K3429006	Promoter	P_degQpromoter	192
BBa_K3429007	Terminator	degQ terminator	36
BBa_K3429011	Coding Sequence	Laccase CotA	1542
BBa_K3429012	Coding Sequence	Blue copper oxidase CueO	1587

Composite Parts

For in vitro characterization

For purification and in vitro characterization of our enzymes we designed the following Composite Parts : cotA, cueO, and ereB. All of these Composite Parts contain a strep-tag for purification and are optimized for E. coli.

For Immobilization on our biofilm

The following parts were designed for immobilization of our enzymes on the TasA matrix protein of our B. subtilis biofilm. In these composite parts the enzymes are genetically fused to TasA. We modeled the structure of the TasA-EreB fusion protein via MD simulation to see whether the enzyme remains stable (BBa_K3429013).

Figure 2: Composite parts for immobilization on biofilm matrix

For our kill switch

Hier ne Abbildung?

Registry Parts

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Part(BBa_)	Type	Description	Length in bp	Part created by
BBa_K733002	Promoter	P_XylA promoter	1387	iGEM HKUST Hong Kong 2012
BBa_K823003	Promoter	P_veg promoter	237	iGEM LMU Munich 2012
BBa_K1680007	Coding Sequence	Cre recombinase	1029	iGEM Tuebingen 2015
BBa_I718016	Recombination Site	lox66 site	34	iGEM Paris 2007
BBa_I718017	Recombination Site	lox71 site	34	iGEM Paris 2007
BBa_K1159000	Coding Sequence	Erythromycin Esterase Type II (EreB) in RFC[25]	1254	iGEM TU Munich 2013