See our bricks below!
Ferulic acid decarboxylase Ferulic acid decarboxylase (FDC) catalyzes the conversion of trans-cinnamic acid to styrene. We isolated this genetic part from Saccharomyces cerevisiae and inserted the protein-coding sequence into the pSB1C3 backbone. The original sequence in yeast contains an SpeI restriction site in the 999th nucleotide position. Thus, we performed site-directed mutagenesis in order to make our part BioBrick compatible. Gene sequencing analysis confirmed that our site-directed mutagenesis was successful.
Ferulic Acid Decarboxylase Ferulic Acid Decarboxylase is used to synthesize styrene from trans-cinnamic acid
Ferulic Acid Decarboxylase with T7 promoter Ferulic acid decarboxylase (FDC) catalyzes the conversion of trans-cinnamic acid to styrene. We isolated this genetic part from Saccharomyces cerevisiae and inserted the protein-coding sequence into the pSB1C3 backbone. The original sequence in yeast contains an SpeI restriction site in the 999th nucleotide position. Thus, we performed site-directed mutagenesis in order to make our part BioBrick compatible. Gene sequencing analysis confirmed that our site-directed mutagenesis was successful. This part includes a T7 promoter, allowing for inducible expression.
Codon-optimized Ferulic Acid Decarboxylase Ferulic acid decarboxylase (FDC) catalyzes the conversion of trans-cinnamic acid to styrene. We codon-optimized the FDC gene from Saccharomyces cerevisiae for expression in E. coli. The decision to use FDC from S. cerevisiae was based on prior work in styrene biosynthesis, notably McKenna (2012). Our construct includes the FDC coding sequence, a T7 inducible promoter, a ribosome binding site, and a FLAG-tag peptide sequence for easy and efficient protein purification. We have sequenced our construct and verified that all these components are indeed present.
Ferulic acid decarboxylase Ferulic Acid Decarboxylase is used to synthesize styrene from trans-cinnamic acid
UbiX with T7 promoter and FLAG tag UbiX is a flavin prenyltransferase that normally plays a role in ubiquinone biosynthesis in E. coli. UbiX transfers a prenyl group from dimethylallyl monophosphate (DMAP) to flavin mononucleotide (FMN), thereby creating a cofactor that happens to be essential to the functionality of FDC. Our genetic construct includes the FDC gene, a T7 inducible promoter, a ribosome binding site, and a FLAG-tag peptide sequence for easy and efficient protein purification. We have sequenced our construct and verified that all these components are indeed present.
Styrene Synthesis Operon Combo Plasmid containing a fusion protein of all three enzymes required to synthesise styrene from the starting organic reagent L-Phenylalanine.
S. aureus type II PanK (CoaA) Type II pantothenate kinase from Staphylococcus aureus, not sensitive to feedback inhibition from CoA.
Pank + hybrid phaCAB + lysis Construct that was cloned to have P(3HB) producing E. coli cells lyse when induced with Arabinose. This would greatly facilitate P(3HB) extraction. We are currently still testing and characterizing this part.
Ptet + Luxl Construct that was cloned to have P(3HB) producing E. coli cells lyse after reaching a certain population density using a quorum sensing promoter. This would greatly facilitate P(3HB) extraction. We are currently still testing and characterizing this part.
Autolysis Construct that was cloned to have P(3HB) producing E. coli cells lyse after reaching a certain population density using a quorum sensing promoter. The difference between this brick BBa_K1692023 is that the basic parts composing this gene are in a different order, which is part of our testing to optimize this system. This would greatly facilitate P(3HB) extraction. We are currently still testing and characterizing this part.
cotz-aeBlue-CipA a fusion protein consisting of a spore coat protein, cotZ (building off work done on Sporobeads by the LMU Munich 2012 iGEM team), and a cellulose binding domain (CipA). Additionally, we decided to add aeBlue, a chromogenic protein, between cotZ and CipA to be able to see with the naked eye whether Bacillus is in a vegetative or a spore state.
CipA Cellulose binding domain adapted from Imperial 2014 iGEM team. Last part of our composite part BBa_BBa_K1692028. We removed the illegal EcoR1 site that the Imperial 2014 iGEM team had in their construc.
RFP gDNA This gene provides the intermediate for synthesizing gRNA targeting RFP (BBa_J04450). When used in conjuction with the CRISPR/Cas system, this gRNA will bind to and digest the RFP plasmid.
amilGFP yellow chromoprotein, RBS and promoter This plasmid contains the amilGFP yellow chromoprotein gene from part BBa_K1033931 with the RBS and promoter from part BBa_K608002.
meffBlue blue chromoprotein with RBS and promoter This plasmid contains the meffBlue blue chromoprotein gene found in the BBa_K1033902 part, with the RBS and promoter from part BBa_K608002.
amilCP blue chromoprotein with RBS and promoter This plasmid contains the amilCP blue chromoprotein gene found in part BBa_K592009 and the RBS and promoter from part BBa_K608002.