## How sequence complexity is calculated

The sequence complexity of an unaligned end is calculated as the product of 'the observed vocabulary-usages' divided by 'the maximal possible vocabulary-usages', for word sizes from one to seven. When multiple breakpoints are used to construct a structural variant, the complexity is calculated as the product of the individual sequence complexities of the breakpoints constituting the structural variant.

The observed vocabulary usage for word size, k, for a given sequence is the number of different "words" of size k that exist in that sequence. The maximal possible vocabulary usage for word size k for a given sequence is the maximal number of different words of size k that can possibly be observed in a sequence of a given length. For DNA sequences, the set of all possible letters in such words is four, that is, there are four letters that represent the possible nucleotides: A, C, G and T. The calculation is most easily described using an example.

Consider the sequence CAGTACAG. In this sequence we observe:

• 4 different words of size 1 ('A,', 'C', 'G' and 'T').
• 5 different words of size 2 ('CA', 'AG', 'GT', 'TA' and 'AC') Note that 'CA' and 'AG' are found twice in this sequence.
• 5 different words of size 3 ('CAG', 'AGT', 'GTA', 'TAC' and 'ACA') Note that 'CAG' is found twice in this sequence.
• 5 different words of size 4 ('CAGT', 'AGTA', 'GTAC', 'TACA' and 'ACAG')
• 4 different words of size 5 ('CAGTA', 'AGTAC' , 'GTACA' and 'TACAG' )
• 3 different words of size 6 ('CAGTAC', 'AGTACA' and 'GTACAG')
• 2 different words of of size 7 ('CAGTACA' and 'AGTACAG' )

Note that we only do the calculations for word sizes up to 7, even when the unaligned end is longer than this.

Now we consider the maximal possible number of words we could observe in a DNA sequence of this length, again restricting our considerations to word of size of 7.

• Word size of 1: The maximum number of different letters possible here is 4, the single characters, A, G, C and T. There are 8 positions in our example sequence, but there are only 4 possible unique nucleotides.
• Word size of 2: The maximum number of different words possible here is 7. For DNA generally, there is a total of 16 different dinucleotides (4*4). For a sequence of length 8, we can have a total of 7 dinucleotides, so with 16 possibilities, the dinucleotides at each of our 7 positions could be unique.
• Word size of 3: The maximum number of different words possible here is 6. For DNA generally, there is a total of 64 different dinucleotides (4*4*4). For a sequence of length 8, we can have a total of 6 trinucleotides, so with 64 possibilities, the trinucleotides at each of our 6 positions could be unique.
• Word size of 4: The maximum number of different words possible here is 5. For DNA generally, there is a total of 256 different dinucleotides (4*4*4*4). For a sequence of length 8, we can have a total of 5 quatronucleotides, so with 256 possibilities, the quatronucleotides at each of our 5 positions could be unique.

We then continue, using the logic above, to calculate a maximum possible number of words for a word size of 5 being 4, a maximum possible number of words for a word size of 6 being 3, and a maximum possible number of words for a word size of 7 being 2.

Now we can compute the complexity for this 7 nucleotide sequence by taking the number of different words we observe for each word size from 1 to 7 nucleotides and dividing them by the maximum possible number of words for each word size from 1 to 7. Here that gives us:

(4/4)(5/7)(5/6)(5/5)(4/4)(3/3)(2/2) = 0.595

As an extreme example of a sequence of low complexity, consider the 7 base sequence AAAAAAA. Here, we would get the complexity:

(1/4)(1/6)(1/5)(1/4)(1/3)(1/2)(1/1) = 0.000347