1) dropped energy difference as much as possible.
2) removed Cs and then Us inside loops as long as energy diff. decreases.
3) Clean up red "error" locations if energy diff. decreases.
4) Drop energy diff only if absolute energy increases.
5) Drop energy diff only if absolute energy decreases.
My ideal solution would be eliminating almost all misfolds by changing only one NT.
If I cannot progress after a while, I may also progress to (2) looking at target loop bases that are mismatched in the natural form. I then try mutations of these bases to break the mismatch pairs. The same criteria apply: mutations that (a) reduce existing mismatches, (b) do not seem to introduce any significant new mismatches, and (c) reduce the delta Free Energy.
I will also start from existing designs by other players, try undoing loop base mods, or mods that make minimal/no dFE gains, and then add mods of my own.
But how do we make a more stable single purpose ribo?
A corollary strategy is after combining, see if you can find low delta/mutation combinations to eliminate - sometimes these become available.
1) get the energy difference as low as possible using the entirety of the allowed number of changes.
2) add in original NTs if it lowers the energy difference, to allow more changes to be made elsewhere.
3) then use up the free change slots again making sure each one lowers the energy difference.
Eventually I hit a wall and then I just start again knowing that the next sequence of changes will be random again and hopefully takes me down a different path to a lower energy difference.
Often large energy differences can be made in the purple substructure region, but because they are "neutral" it might be acceptable to lock those off with strong pairs and ignore the change counter and ignore the energy difference increase.
1) Using all the allowed "change" slots, get the energy difference as low as possible manually. e.g. 18/18 changes.
2) Go over the 18/18 limit by 1. e.g. 19/18 changes, making sure the energy difference is still lower than the 18/18 string.
3) Load in the script that jandersonlee provided in this post ("Mark Mutations") that automatically uses the mark tool to mark modifications to the original puzzle structure.
4) Then run the already provided in-game Mutation/Submission tool that will run through individual permutations of all marked bases.
5) Using the z + y (undo + redo) buttons on keyboard, search through the permutations until you find one that has 18/18 change slots filled and is a lower energy than the original 18/18 in step 1.
6) Submit solution.
7) This new string is unique from the original 18/18 string in step 1 and the 19/18 string in step 2, so you can run another Mutation/Submission task for free to see if you can find a lower energy 17/18 or 18/18 string. If you can't just go back and repeat step 1.
So far I have found this to be a good evolution driven approach to lowering the energy difference.
Brourd mentioned that our 23S designs may not have performed as well as the 16S designs because the 23S subunit is the catalytic center. We also know that one of the best 16S designs placed mutations in only one region of the 16S, specifically in one hairpin. Therefore I suggest we test designs that make mutations in only one domain of the 23S.
There are six regions to the 23S and they largely fold independently. We probably should avoid or be extremely careful in domain 5, the home of the peptidyl transferase center (PTC) where the Big Action happens. I’m going to put a hashtag in my design title indicating which domain and helix I am altering, such as #D1 #H18 #H21. Players using a different design approach may still want to designate the domain or helices in the design title for quicker data analysis. (Do we want to use Roman numerals in our hashtag? I’m not a Roman numeral fan.)