You already wrote two for loops in your list comprehension, you can just reuse them.
(I modified your list comprehension, I used y and x instead of 2 x)
seq1 = 'asdfasdfasdf'
seq2 = 'asdfasdfasdf'
mymatrix = [[0 for y in range(len(seq1)+1)]for x in range(len(seq2)+1)]
# ^^^^^^^^^^^^^^^^^^^^^^^^^^ reuse this
d = {'score' : 0, 'pointer' : 'none'}
for y in range(len(seq1)+1):
mymatrix[0][y] = d.copy()
# The first one is already covered, just leave it off using slice
for x in range(len(seq2)+1)[1:]:
mymatrix[x][0] = d.copy()
for line in mymatrix:
print(line)
Out:
[{'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}, {'pointer': 'none', 'score': 0}]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]
[{'pointer': 'none', 'score': 0}, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0]