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409_C_1 (or -6C) ‏

409_C_1 (or -6C) ‏. 409_A. 409_C. 409_G (WT) ‏. 409_T. a: -801.51 c: -863.91 g: -857.45 t: -859.32. 407_G_1 (or -8G) ‏. 407_C. 407_G. 407_T. 407_A (WT) ‏. g: -876.05 a: -870.01 c: -870.33 t: -869.29. 422_C_1 (or +8C) ‏. 422_C. 422_G. 422_T. 422_A (WT) ‏.

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409_C_1 (or -6C) ‏

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  1. 409_C_1 (or -6C)‏ 409_A 409_C 409_G (WT)‏ 409_T a: -801.51 c: -863.91 g: -857.45 t: -859.32

  2. 407_G_1 (or -8G)‏ 407_C 407_G 407_T 407_A (WT)‏ g: -876.05 a: -870.01 c: -870.33 t: -869.29

  3. 422_C_1 (or +8C)‏ 422_C 422_G 422_T 422_A (WT)‏ c: -849.70 g: -838.41 a: -835.20 t: -838.70

  4. (-) TGAGGAGGTT TCTCTGTAAG (+)‏ Design accessed intended position, wt didn’t originally cut it Designs that make protein nonspecific at position WT doesn’t really care -10, -5, +5, +8, +9, +10 -9C, -8T, -8G, -6C, -3 (T), +3 (A)‏ Most important remaining Some off-target activity from designs Design made more specific +5C, +8C, +8A -9T, -9G, -7, -6A, -4C, -4T, +4, +6, +7C, +7G -6T, -4A, +7A

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