Unaligned, compressed, raw movie frames of AAV2-L336C. The gain normalization reference images (norm) are included. No dark gain reference images are present because dark images values are always zero in counting mode camera. Magnification anisotropy measured using gold diffraction was found to be 1.005 at major axis, 0.995 at minor axis, at 7.7 degrees angle. Total dose was 22.5 electrons per Å squared over 3.5 seconds. Calibrated pixel size is 0.394 Å/pixel.

MotionCor2 aligned, gain corrected, magnification anisotropy corrected, Fourier bin by 2 movie frames of AAV2-L336C. Movie frames were aligned using MotionCor2 with 5 by 5 patches, a grouping of 3 and B-factor of 100, and Fourier space binning of 2 (resulting in a pixel size of 0.788 Å/pixel) through the Appion software package. No dose weighting was applied for these movies frames.

MotionCor2 aligned, gain corrected, magnification anisotropy corrected, Fourier bin by 2 movie frames of AAV2-L336C. Movie frames were aligned using MotionCor2 with 5 by 5 patches, a grouping of 3 and B-factor of 100, and Fourier space binning of 2 (resulting in a pixel size of 0.788 Å/pixel) through the Appion software package. Dose weighting was applied for these summed micrographs, using a dose per frame of 0.3214 electrons per Å squared.

MotionCor2 aligned, gain corrected, magnification anisotropy corrected, Fourier bin by 2 movie frames of AAV2-L336C. Movie frames were aligned using MotionCor2 with 5 by 5 patches, a grouping of 3 and B-factor of 100, and Fourier space binning of 2 (resulting in a pixel size of 0.788 Å/pixel) through the Appion software package. These are the non-dose weighted summed micrographs that can be used for CTF estimation.

MotionCor2 aligned, gain corrected, magnification anisotropy corrected, Fourier bin by 2 movie frames of AAV2-L336C. Movie frames were aligned using MotionCor2 with 5 by 5 patches, a grouping of 3 and B-factor of 100, and Fourier space binning of 2 (resulting in a pixel size of 0.788 Å/pixel) through the Appion software package. Frames 5 to 19 were summed and no dose weighting was applied, as these corresponded to frames that gave the best resolution reconstructions individually compared to the other frames. Note that the signal-to-noise ratio in these micrographs are less those from all 70 frames, and should only be used for the final refinements and reconstructions once Euler angles and shifts have been established.

Final set of AAV2-L336C particles extracted from MotionCor2 aligned, gain corrected, magnification anisotropy corrected, Fourier bin by 2, not dose weighted, frames 5 to 19 summed micrographs. A Relion .star format file is provided and contains the final Euler angles, shifts and also the micrograph coordinates.

From the final set of 30515 particles, each particle was expanded into 3: a particle for frames 5-9, one for frames 10-14 and one for frames 15-19. This resulted in a stack 3 times larger that was used for rotational alignment in Frealign. A .mrc stack and Frealign .par file are provided with the final Euler angles and shifts.