Improved PsacB promoter

We had made improvement on part BBa_K4043004. The original parts was the endogenous promoter of B. subtilis that can respond to the sucrose concentration in the medium. We replaced its constitutive promoter with the Pveg promoter and repeat the regulatory sequence for 1, 2, 3，and 5 times. By changing the constitutive promoter and multiplying the regulatory sequence , we are able to improve the degree of induction and make the induction more stable at most growth conditions (although maximally during exponential growth). You can find more information about our brick in the Registry pages of BBa_K4202025, BBa_K4202026, BBa_K4202027, and BBa_K4202028.

Vector

BBa_K4202016 is the shuttle vector(pHY-300-PLK) used in our project.

Calcium Carbonate Production Module

BBa_K4202004, a codon-optimized sequence encoding carbonic anhydrase, was used to enhance the expression of carbonic anhydrase in Bacillus subtilis to enhance the production of calcium carbonate and biomineralization. However, the characterization results showed that the new part did not enhance the biomineralization effect compared to the original part (BBa_K2232000), but rather weakened the effect.

BBa_K4202008 is the coding sequence of Amorphous Calcium Carbonate Binding Protein(ACCBP) with signal peptide and BBa_K4202009 is the coding sequence of ACCBP without signal peptide. BBa_K4202010 encoded the shorter version of ACCBP with the absence of 16 amino acids at N terminus.

Biological scaffold Module

BBa_K4202002 and BBa_K4202003 encoded mRFP:SpyTag fusion protein and mRFP:SpyCatcher fusion protein, which were used to verify the combination of SpyTag and SpyCatcher. BBa_K4202006 and BBa_K4202015 are proteins used in the bio-scafford which encoded the Hag:SpyTag and EutM:SpyCatcher fusion protein, respectively.

Quorum Sensing Module

BBa_K4202007 is a composite part, consisting of a oxygen-limitation induced promoter, RBS, and a gene cluster composed of spaBCTS. This part is used to induce the expression of subtilisin in oxygen-limitation environment deep in the crack to activate the expression of genes downstream of the chymotrypsin-inducible promoter.

Biosafety Module

BBa_K4202045 is the coding sequence of our kill switch used in the suicide of our engineered bacteria.BBa_K4202001 and BBa_K4202046 are codon optimized sequence that coding mazF-mazF toxin-antitoxin system. They worked together in the kill switch of our engineered bacteria. BBa_K4202019 and BBa_K4202020 are the basic parts involved in the construction of the engineered sucrose-sensitive promoter. BBa_K4202025, BBa_K4202026, BBa_K4202027, and BBa_K4202028 are our engineered sucrose-sensitive promoters which were more sensitive to the low concentration of sucrose. Other parts are the generator we used to characterize the original part (BBa_K4202029, BBa_K4202030, BBa_K4202031, and BBa_K4202032) and the improved part (BBa_K4202033, BBa_K4202034, BBa_K4202035, BBa_K4202036, BBa_K4202037, BBa_K4202038, BBa_K4202039, BBa_K4202040, BBa_K4202041, BBa_K4202042, BBa_K4202043, and BBa_K4202044).