Decide on an overall cloning strategy for the project

So biology also needs to finalize a decision between:

1) Cloning in those growth slowers under a number of the Anderson promoters (http://parts.igem.org/Promoters/Catalog/Anderson) — this will require picking a subset of these promoters spread between 0 and 1 in strength and then transforming them from last year's iGEM kit plates! Because these are already characterized in terms of their strength, you just need to clone them in front of the growth slowers! That means P x G plasmids, where P is the number of promoters you pick and G is the number of growth slowers. 2) Alternatively, you could clone the growth slowers downstream from an inducible promoter that's "highly-titratable". This means G + 1 plasmids will be needed, since now everything uses the same promoter, but you'll need to clone some reporter gene (with a high dynamic range) downstream of the promoter as well (that's the +1) so that you can calculate how much expression results from a given amount of inducer.

Overall, option 1 means cloning a lot more plasmids (but this is something you can do, theoretically, with golden gate cloning) but it results in super simple experiments (just a growth curve). Option 2 is way less cloning, but means that every growth curve will have to be done with inducer present and you'll need to pick a reliable reporter gene to build a "standard curve" with since you now no longer know how much the genes are expressed relative to other inducer concentrations. Additionally, you actually need to find that inducible promoter and reporter gene, whereas the Anderson promoters we should have already!

Once you decide which approach to take, you can design and order primers!

Sheffield-iGEM / .github

Decide on an overall cloning strategy for the project #3