Our plasmid is composed of three parts: a promoter and a terminator Biobrick as well as a novel addition to the biobrick library, the phsABC gene that is known to encode Thiosulfate Reductase.(1) phsABC gene and vector
This central component encodes Thiosulfate Reductase. The gene phsABC was obtained through Addgene from Dr. Jay Keasling's laboratory at University of California, Berkeley. According to their results, thiosulfate reductase encoded in the plasmid pSB74 showed the highest activity catalytic activity, so we obtained phsABC from the plasmid pSB74.
Promoter used was designed by Caitlin Conboy and was found within the parts registry. This promoter is a Quad Part Inverter: “that is, a PoPS-based inverter composed of four sub-parts: a ribosome binding site, a coding region for a repressor protein (e.g., lambda cI), a terminator, and the promoter (e.g., pLambda) regulated by the encoded repressor protein.” Research into promoter activity by previous groups has suggested that this promoter has a strong on state with a noticeable background in the off state .
Length 1370 bp
(3) Biobrick Terminator The terminator used was the 129 bp BBa_B0015 designed by Reshma Shetty. It is actually a double terminator composed of BBa_B0010 and BBa_B0012 and the BioBrick assembly scar & was chosen for its reliability and availability.
Restriction Enzyme Sites:
The digested sticky ends of the enzymes Xba I and Spe I are complimentary. Once two ends from different combine, neither Xba I nor Spe I can recognize its restriction site in the gene.
By using this ligation method, inserted phsABC into B0015:
Read more about the background of the phsABC gene