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Quantification of HaloTag labels inside StrepEncSM by single‐molecule fluorescence. A) smFRET measurements of StrepEncSM{HaloTag*JF549*JF646}. All photon burst search (APBS), time window 500 μs, threshold 50 photons, FRET‐2CDE filter: 0–10, DA rate: 15–20 kHz, DD rate: 2–20 kHz, burst duration 3–20 ms. B) Dual color burst search (DCBS), time window 500 μs, DD rate: 2–200 kHz, AA rate 5–50 kHz, DA rate: 20–150 kHz, FRET‐2CDE filter: 5–15, burst duration 2–7 ms, 2 Gaussian fits.
