Team:Linkoping Sweden/Partscomposite
Assembly of multiple basic parts results in a composite part. Alternative combinations with these parts can create various larger circuits with different functions. These can then be transformed into living organisms.
Apart from the improved BioBrick, all the composite parts consist of Linker-GFP (BBa_K3892005) assembled with one of our coding basic parts. This assembly is enabled by a BamHI cleaving site before the linker and after the gene in the coding parts as seen in Figure 1. The potential use of the resulting composite parts (BBa_K3892011-BBa_K3892015) is further described on our design page.
Figure 1. Photos taken from Benchling to illustrate how our basic coding parts can be assembled with part BBa_K3892005. A) Displays cleaving with BamHI restriction enzyme of our halorhodopsin part as well as the Linker-GFP part. B) Illustrates the resulting composite part BBa_K3892011 obtained after ligation of the cleaved parts in A).
The improved version of the BioBrick BBa_K1073022 is listed below as EforRed improved (BBa_K3892016). It was successfully improved by adding the G-10 leader BBa_K3892010. Figure 2. illustrates the differences between the two parts and the results obtained from the improved part can be further read on our improvement page.
Figure 2. Photos taken from Benchling. A) Shows the part we aimed to improve. The light blue annotations indicate the parts that were exchanged in the improved version, meaning a weak RBS separated by two spacer regions. B) Shows the improved part we designed. The light blue annotations indicate the improved regions, meaning a G-10 leader containing a strong RBS separated by two spacer regions.
Table 1. All composite parts 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_K3892011 | Beta-carotene 15,15'-dioxygenase-Linker-GFP | Beta-carotene splitting enzyme for all-trans-retinal production with attached Green fluorescent protein | 1883 |
| BBa_K3892012 | Halorhodopsin-Linker-GFP | Anion pump for inward transport of chloride ions with attached Green fluorescent protein | 1703 |
| BBa_K3892013 | Slr1272-CBD-Linker-GFP | Native slr1272 protein derived from Synechocystis sp. PCC 6803 and optimized carbohydrate-binding domain with a C-terminal linker (CBDcipA) and Green fluorescent protein for separation of bacteria from water | 2084 |
| BBa_K3892014 | MscL-Linker-GFP | Mechanosensitive cation channel for outward transport of ions with attached Green fluorescent protein | 1265 |
| BBa_K3892015 | Channelrhodopsin-Linker-GFP | Cation channel derived from Chlamydomonas reinhardtii for inward transport of sodium ions | 3041 |
| BBa_K3892016 | EforRed improved | Red chromoprotein with strong RBS | 758 |