Team:Open Science Global/Parts

Part Collection


The main goal of our project is to make a frugal biofoundry capable of producing high quantities of enzymes and other proteins of interest. To do that, our team designed a library of parts in order to test in a high-throughput way the combinations that would generate the best results. For this year, the most important part generated by our team was the pHT43 plasmid (BBa_K4103000), an improvement on (BBa_K1074001) from 2013 - this plasmid acts as a shuttle vector between E. coli and B. subtilis, and allows the efficient production and secretion of our protein-of-interest, with our team focusing efforts to use this vector for the production of Pfu-Sso7d Polymerase (BBa_K4103011) codon optimized for B. Subtilis using the software created by our team.

For how we made our parts, see "Friendzymes BioBlaze Notebook" and "Friendzymes OSN Notebook" (too large to embed).

Basic Parts

Part Name Type Description Length (bp) Conjugation origin Transfer Origin pIP501 319 Resistance marker kanR 968 Resistance marker SpecR 990 Origin of replication EcoOri 589 Resistance Marker catA9 946 Ribozyme Insulator SarJ 79 Insulator HDV-1 (3' Ribozyme Insulator) 85 Vector pOpen_v3 2687 Assembly Connector ACLstart 191 Assembly Connector ACRend 170 Toxin/Negative Selection Txn_ccdB 933 Homology Arm 5'lacA 508 Homology Arm 3'lacA 408 Homology Arm 5'AmyE promoter 521 Homology Arm 3'AmyE 498 Homology Arm 5'ThrC 501 Homology Arm 3'ThrC 429

Composite Parts

Part Name Type Description Length (bp) Plasmid pHT43_Pfu_sso7d 9484 Promoter + Insulator lacI+riboJ 1614 RBS_SecTag AmyQ 135 CDS Pfu_sso7d_2 2534 5' Tags TEV-GFP-10xHis_R5 C-Tag 865 Library Plasmid SecTag Library 1 6968 Library Plasmid SecTag Library 2 4962 Library Plasmid SecTag Library 3 4968 Library Plasmid SecTag Library 4 4879 Library Plasmid SecTag Library 5 4913 Library Plasmid SecTag Library 6 4868