yes, good points - this is true of any flat image, dslr or not. precise focus is probably not super important, but you should not be wildly out of focus. most important is the camera angle, as the shape of the vignetting will change with the angle and any dust motes that are on optical surfaces in front of the sensor will move as the angle changes. there's nothing worse than a dust donut that's just slightly off - it ends up looking "embossed" on the calibrated image...
so if you've removed the camera from the OTA then it's probably better to just take flats next time rather than trying to replicate your camera angle.
with the lightbox be careful that your flat exposure is long enough to average out any flickering in your light source. for instance a fluorescent light has a 60hz frequency so short flat exposures will not be evenly illuminated. if you have to lower the ISO, that's okay, just calibrate the flat subs with bias frames (or dark flats) taken at the same ISO.
rob