Team:Linkoping Sweden/Part Collection
This part collection serves as a complete tool for assembly, expression and desalination of seawater by Synechocystis sp. PCC 6803 followed by separation of the bacteria from the desalinated water. Each part in the collection has been designed to perform its individual task as a part of our bigger desalination system. All parts were designed to function on their own as well as together with each other. Our part collection would function where halorhodopsin, channelrhodopsin, and beta-carotene 15,15'-dioxygenase would produce the central desalination mechanism. MscL would improve the fitness of the cells and prevent cell disruption. The Slr1272-CBD would enable a cheap and easy way to separate our modified bacteria from the water. In combination, all these parts would create a desalination system with a safe separation system to ensure that no modified bacteria is released into the wild. The parts BBa_K3892000, BBa_K3892001, BBa_K3892002, BBa_K3892004, and BBa_K3892007, further described in Table 1, contain a BamHI restriction enzyme cleaving site on their 3’ end. This makes them compatible to assemble with the part BBa_K3892005, as illustrated in Figure 1. These resulting, assembled, parts are featured on our composite parts page. The potential implementation of our basic parts can be read further on our design page and all our basic parts can be found on our basic parts page.
Figure 1. A schematic picture illustrating how our Linker-GFP part can be assembled together with our coding basic parts to create our composite parts by their 3’ terminal BamHI restriction enzyme cleaving site.
The part BBa_K3892008 is a native, strong constitutive promotor derived from Synechocystis sp. PCC 6803. It would be used for all the parts involved in desalination and separation. The part BBa_K3892009 contains a spacer region, followed by a ribosome binding site (BBa_B0034) and then another spacer. This would also be used for all the desalination and separation parts. Similar to the promoter, this RBS is strong in both E. coli and Synechocystis sp. PCC 6803. The reason for using a strong protomer in combination with a strong RBS is to achieve high expression to ensure a sufficient amount of halorhodopsin, channelrhodopsin, beta-carotene 15,15'-dioxygenase, MscL and slr1272-linker-CBD.
Table 1. Table displaying the parts that were central to our project.
| Biobrick ID | Short name | Short description | Length |
|---|---|---|---|
| BBa_K3892000 | Beta-carotene 15,15'-dioxygenase | All-trans retinal generating enzyme from Natronomonas pharaonis | 1059 |
| BBa_K3892001 | Halorhodopsin | Light-activated anion pump derived from Natronomonas pharaonis | 879 |
| BBa_K3892002 | Linker-CBD | Carbohydrate binding domain with a C-terminal linker (CBDcipA) derived from Clostridium thermocellum | 498 |
| BBa_K3892003 | Slr1272 | S-layer protein derived from Synechocystis sp. PCC 6803 | 762 |
| BBa_K3892004 | MscL | Mechanosensitive ion channel from Synechocystis sp. PCC 6803 | 441 |
| BBa_K3892005 | Linker-GFP | Green fluorescence protein from Aequorea victoria | 732 |
| BBa_K3892007 | Channelrhodopsin | Light-activated cation pump derived from Chlamydomonas reinhardtii | 2217 |
| BBa_K3892008 | PpsbA2 | Native, strong constitutive promotor derived from Synechocystis sp. PCC 6803 | 38 |
| BBa_K3892009 | Spacer-RBS-Spacer | Ribosome binding site with spacers | 45 |