Parts

Basic Parts

Type Part Name Short Description Base Pair
BBa_25G5YR4C pMA5 plasmid backbone The pMA5 plasmid backbone is used as the primary vector for our final plasmid pMA5-pls-PPK2-lac-alrA plasmid 11117bp
BBa_25EMGU3Q pls pls is the gene responsible for the production of ε-Polylysine 3960 bp
BBa_25L5WCKD PPK2 PPK2 is the gene responsible for ATP regeneration that can provide our strain with more energy 963 bp
BBa_252TM36G lysC311 lysC311 is used to subsitute the thrD gene in order to relieve the lysine inhibition 1266 bp
BBa_25IHX7XE thrD-Left thrD-Left is connected to the lysC311 to form a plasmid able to insert into the Bacillus subtilis 309 bp
BBa_25F7CW8V thrD-Right thrD-Right is also connected to the lysC311 to form a plasmid able to insert into the Bacillus subtilis 299 bp
BBa_25XEU57S sg thrD sg thrD help guide the Cas 9 protein to knock off the thrD gene in Bacillus subtilis through CRISPER/Cas 9 meditated homologous recombination replacement 20 bp
BBa_2571HMVC sg alrA sg alrA helps guide CAs 9 protein to knock off the alrA gene in the Bacillussubtilis through the Cas9-sg RNA system and thus help ensure the biosafety of our engineer bacteria 20 bp
BBa_25XOTKEA Lac operon The lac operon is used as a regulatory control unit in this project and it helps us to complete our suicide switch design 2385 bp
BBa_25KA4J1C pJMP3 plasmid backbone pJMP3 is the plasmid backbone used to help insert sg RNA into our targeted microorganism to help the Cas 9 protein successfully knock off intended DNA sequences. 7197bp
BBa_25ESL37E PTH43 plasmid backbone PHT43 is the original vector for the lac operon when we obtain it and we used it as a base for later translation of the lac-alrA unit into pMA5-pls-PPK2 plasmid 8056 bp
BBa_25QQN8OK PKY300PKL plasmid backbone the PKY300PKL plasmid backbone is the vector of our thrD-lysC311 plasmid in the second cycle used to complete replacement of thrD gene with lysC311 gene to relieve lysine inhibition in Bacillus subtilis 7571 bp
BBa_251M08GX alrA alrA is the gene that's responsible for encoding alanine racemase in Bacillus subtilis naturally. In this project, we knock off the alrA in bacillus subtilis for our design of a killer switch 1170 bp

Composite Parts

Registry Code Part Name Short Description Base Pair
BBa_25EJCCFR pMA5-pls This composite part is used to help insert the pls gene into Bacillus subtilis. 11117 bp
BBa_25404K7H pMA5-pls-PPK2 This composite part is responsible of the insertion of pls and PPK2 gene. 12080 bp
BBa_25TI2N93 pJMP3-sgthrD This part is the plasmid in which the sgthrD is on. It help the insertion sg thrD and thus the deletion of thrD gene in Bacillus subtilis. 7197 bp
BBa_25QWQUAM PHY300PLK-thrDL-lysC311-thrDR This plasmid is used to transfer the lysC311 gene into Bacillus subtilis, thus completeing the substitution of thrD with lysC311 7571 bp
BBa_25BTTP42 pMA5-pls-PPK2-lac-arlA The pMA5-pls-PPK2-lac-alrA plasmid is a comprehensive plasmid designed to optimize ε-polylysine production and ensure biosafety control in Bacillus subtilis. This plasmid integrates several crucial genetic elements that work together to achieve both efficient polylysine biosynthesis and regulated bacterial growth, making it a highly specialized tool for biotechnology applications. 13827bp
PHY300PLK-Cas9 PHY300PLK-thrDL-lysC311-thrDR Plasmid This plamid help insert the Cas 9 protein crucial for gene knock off in Bacillus subtilis using the CRISPER meditated homologous recombination replacement. 10729 bp

Biocontainment Parts

Registry Code Part Name Short Description Base Pair
BBa_259DQLVW PJMP3-sg alrA This part is used to insert sg alrA into our engineered bacteria so that alrA could be knocked off in Bacillus subtilis 7197 bp
BBa_25KEDONE PTH43-lac-alrA This part is used as a base for the modification and creation of our final pMA5-pls-PPK2-la-alrA plasmid. 9226 bp