Team:Linkoping Sweden/Partsbasic
BioBricks are the key elements of synthetic biology. They are the shortest DNA sequences that encode a unit with a specific biological function. It could be a coding sequence, a promoter or other. In our project, we designed 11 basic parts among which eight are coding and central to our project. The parts BBa_K3892000, BBa_K3892001, BBa_K3892002, BBa_K3892004 and BBa_K3892007 contain a BamHI restriction enzyme cleaving site on their 3’ end, making them compatible to assemble with a part BBa_K3892005, illustrated in Figure 1. The potential implementation of our basic parts can be read further on our design 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 promoter 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 as well as a strong RBS is that a high expression is wanted, to ensure a sufficient amount of halorhodopsin, channelrhodopsin, beta-carotene 15,15'-dioxygenase, MscL and slr1272-linker-CBD.
BBa_K3892006 and BBa_K3892010 were used to create an improved part. The use of these parts is further explained on our improvement page.
Table 1. All basic parts that were designed for our project with the contribution of all team members. All team members worked with the parts in the wet-lab. The potential uses of these parts are further described on their respective Part Registry page. Press the part number to reach the Part Registry page for that specific BioBrick.
| 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_K3892006 | EforRed | Chromoprotein EforRed from Echinopora forskaliana | 687 |
| BBa_K3892007 | Channelrhodopsin | Light-activated cation channel 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 |
| BBa_K3892010 | G-10 leader and RBS | Based on BBa_K1758100, ribosome binding site; phage T7 gene 10 leader | 36 |