reform is a python-based command line tool that allows for fast, easy and robust editing of reference genome sequence and annotation files.
Execution of reform requires a reference sequence (fasta), reference annotation (GFF or GTF), the novel sequences to be added (fasta), and corresponding novel annotations (GFF or GTF). A user provides as arguments the name of the modified chromosome and either the position at which the novel sequence is inserted, or the upstream and downstream sequences flanking the novel sequences. This results in the addition and/or deletion of sequence from the reference in the modified fasta file. In addition to the novel annotations, any changes to the reference annotations that result from deleted or interrupted sequence are incorporated into the modified gff. Importantly, modified gff and fasta files include a record of the modifications.
Learn more at https://gencore.bio.nyu.edu/reform/
reform requires Python3, pgzip and Biopython v1.78 or higher.
Install pgzip and biopython if you don't already have it:
pip install pgzip
pip install biopython
Invoke the python script:
python3 reform.py
--chrom=<chrom> \
--position=<pos> \
--in_fasta=<in_fasta> \
--in_gff=<in_gff> \
--ref_fasta=<ref_fasta> \
--ref_gff=<ref_gff>
chrom
ID of the chromsome to modify
position
Position in chromosome at which to insert -1
to add to end of chromosome. Note: Either position, or upstream AND downstream sequence must be provided. Note: Position is 0-based
upstream_fasta
Path to Fasta file with upstream sequence. Note: Either position, or upstream AND downstream sequence must be provided.
downstream_fasta
Path to Fasta file with downstream sequence. Note: Either position, or upstream AND downstream sequence must be provided.
in_fasta
Path to new sequence to be inserted into reference genome in fasta format.
in_gff
Path to GFF file describing new fasta sequence to be inserted.
ref_fasta
Path to reference fasta file.
ref_gff
Path to reference gff file.
python3 reform.py
--chrom="I" \
--upstream_fasta="data/up.fa" \
--downstream_fasta="data/down.fa" \
--in_fasta="data/new.fa" \
--in_gff="data/new.gff" \
--ref_fasta="data/Saccharomyces_cerevisiae.R64-1-1.dna.toplevel.fa" \
--ref_gff="data/Saccharomyces_cerevisiae.R64-1-1.34.gff3"
reformed.fa
Modified fasta file.
reformed.gff3
Modified GFF file.