iGEM Stockholm 2018 Wiki

Parts

This page provides an overview of all BioBricks that we have created and submitted to the iGEM Registry of Standard Biological Parts. For more detailed information and characterization of these parts, use the links below to reach the Registry page of each BioBrick.

Creation of Our BioBricks

We used different methods for constructing our parts. The laccase enzyme was based on an already existing BioBrick (BBa_K500002 designed by the Tianjin team 2010), where we removed the native secretion signal and added a His-tag via overhang PCR. The mutated version of this laccase (based on our modeling results) was ordered from IDT, making use of their kind sponsorship and saving valuable time. In our process of improving a BioBrick, we utilized a recently developed site-directed mutagenesis technique. In order to test our improved BioBrick, a functional construct including our BioBrick (RBS + fluorescent protein) and a promoter was assembled by restriction and ligation.

Improved BioBrick

In our project, we explored the possibilities of a site-directed mutagenesis technique called SAMURAI. For this purpose, we chose to mutate an already existing BioBrick, the composite part BBa_I13502 designed by the MIT team 2005. This part encodes a monomeric red fluorescent protein (mRFP1) and includes a ribosome binding site (RBS). SAMURAI was applied to remove two AgeI restriction sites, making the part compatible with RFC25. We named our improved BioBrick BBa_K2835000 (see table below).

All Parts

Name	Type	Description	Designer	Size (bp)
BBa_K2835000	Translational unit	mRFP1 with RBS, compatible with RFC25	Emma Larsson, Hanna Norbäck	724
BBa_K2835001	Composite part, reporter	mRFP1 with constitutive promoter and RBS, compatible with RFC25	Emma Larsson, Hanna Norbäck	767
BBa_K2835007	Composite part, reporter	mRFP1 with constitutive promoter and RBS (not compatible with RFC25, created to compare it with BBa_K2835001)	Emma Larsson, Hanna Norbäck	767
BBa_K2835003	Protein domain	Laccase with His-tag, fungal enzyme from Trametes versicolor	Els Alsema, Marit Möller	1518
BBa_K2835004	Protein domain	Mutated laccase with His-tag, mutated for increased activity towards sulfamethoxazole (antibiotic)	Darko Mitrovic	1518
BBa_K2835005	Basic part, coding	Laccase with His-tag and start codon (ATG), fungal enzyme from Trametes versicolor	Els Alsema, Marit Möller	1521
BBa_K2835006	Basic part, coding	Mutated laccase with His-tag and start codon (ATG), mutated for increased activity towards sulfamethoxazole (antibiotic)	Darko Mitrovic	1521

Basic Parts

Name	Type	Description	Designer	Size (bp)
BBa_K2835000	Translational unit	mRFP1 with RBS, compatible with RFC25	Emma Larsson, Hanna Norbäck	724
BBa_K2835003	Protein domain	Laccase with His-tag, fungal enzyme from Trametes versicolor	Els Alsema, Marit Möller	1518
BBa_K2835004	Protein domain	Mutated laccase with His-tag, mutated for increased activity towards sulfamethoxazole (antibiotic)	Darko Mitrovic	1518
BBa_K2835005	Basic part, coding	Laccase with His-tag and start codon (ATG), fungal enzyme from Trametes versicolor	Els Alsema, Marit Möller	1521
BBa_K2835006	Basic part, coding	Mutated laccase with His-tag and start codon (ATG), mutated for increased activity towards sulfamethoxazole (antibiotic)	Darko Mitrovic	1521

Composite Parts