代码如下：

#!/usr/bin/env python3
# -*- coding: utf-8 -*-
# extract_splice_sites.py
# @author Aaron Liu
# @description 从gtf文件中提取剪切位点（参考hsiat2_extract_splice_sites.py）
# @created 2020-09-04T19:26:12.427Z+08:00
# @last-modified 2020-09-05T17:33:50.935Z+08:00

from argparse import ArgumentParser, FileType

def extract_splice_sites(gtf_file):
    trans = {}
    for line in gtf_file:
        line = line.strip()
        if not line or line.startswith('#'):
            continue
        if '#' in line:
            line = line.strip('#')[0].strip()

        chr, source, feature, start, end, score, strand, \
            frame, values = line.split('\t')
        start, end = int(start), int(end)

        if feature != 'exon' or start >= end:
            continue

        value_attr = {}
        for attr in values.split(';'):
            gene, _, transcript = attr.strip().partition(' ')
            value_attr[gene] = transcript.strip('"')

        if "gene_id" not in value_attr \
            or "transcript_id" not in value_attr:
            continue

        transcript_id = value_attr['transcript_id']
        gene_id = value_attr['gene_id']
        if transcript_id not in trans:
            trans[transcript_id] = [gene_id, chr, strand, [[start, end]]]
        else:
            trans[transcript_id][3].append([start, end])

    for transid, [gene_id, chr, strand, exon] in trans.items():
        exon.sort()
        tem_exon = [exon[0]]
        for i in range(1, len(exon)):
            if exon[i][0] - tem_exon[-1][1] <= 5:
                tem_exon[-1][1] = exon[i][1]
            else:
                tem_exon.append(exon[i])
        trans[transid] = [gene_id, chr, strand, tem_exon]

    junction = set()
    for gene, chrom, strand, exon in trans.values():
        exon.sort()
        for i in range(1, len(exon)):
            junction.add(
                (gene, chrom, exon[i - 1][1] + 1, exon[i][0] - 1, strand))

    junction = sorted(junction)
    return junction


if __name__ == "__main__":
    parse = ArgumentParser(description='Extract splice sites from a GTF file')
    parse.add_argument('-i',
                       '--gtf_file',
                       required=True,
                       nargs='?',
                       type=FileType('r'),
                       help="a gtf file is required")
    parse.add_argument('-o',
                       '--out_file',
                       required=True,
                       nargs='?',
                       type=FileType('w'),
                       help='output file is required')

    args = parse.parse_args()

    splicesites = extract_splice_sites(args.gtf_file)
    for row in splicesites:
        args.out_file.write(row[0] + '\t' + row[1] + '\t' + str(row[2]) +
                            '\t' + str(row[3]) + '\t' + row[4] + '\n')

实例：

./extract_splice_sites.py -i merge.gtf -o ss.txt

输出：

...
MSTRG.10013 chr04   2534745 2535071 +
MSTRG.10013 chr04   2535164 2535818 +
MSTRG.10013 chr04   2535988 2536609 +
MSTRG.10013 chr04   2536732 2537314 +
MSTRG.10013 chr04   2536732 2537381 +
MSTRG.10013 chr04   2537444 2537540 +
MSTRG.10013 chr04   2537444 2537551 +
MSTRG.10013 chr04   2537651 2537730 +
MSTRG.10013 chr04   2537789 2537870 +
...