minimap2/misc/paftools.js

2536 lines
81 KiB
JavaScript
Executable File

#!/usr/bin/env k8
var paftools_version = '2.16-r933-dirty';
/*****************************
***** Library functions *****
*****************************/
/*******************************
* Command line option parsing *
*******************************/
var getopt = function(args, ostr) {
var oli; // option letter list index
if (typeof(getopt.place) == 'undefined')
getopt.ind = 0, getopt.arg = null, getopt.place = -1;
if (getopt.place == -1) { // update scanning pointer
if (getopt.ind >= args.length || args[getopt.ind].charAt(getopt.place = 0) != '-') {
getopt.place = -1;
return null;
}
if (getopt.place + 1 < args[getopt.ind].length && args[getopt.ind].charAt(++getopt.place) == '-') { // found "--"
++getopt.ind;
getopt.place = -1;
return null;
}
}
var optopt = args[getopt.ind].charAt(getopt.place++); // character checked for validity
if (optopt == ':' || (oli = ostr.indexOf(optopt)) < 0) {
if (optopt == '-') return null; // if the user didn't specify '-' as an option, assume it means null.
if (getopt.place < 0) ++getopt.ind;
return '?';
}
if (oli+1 >= ostr.length || ostr.charAt(++oli) != ':') { // don't need argument
getopt.arg = null;
if (getopt.place < 0 || getopt.place >= args[getopt.ind].length) ++getopt.ind, getopt.place = -1;
} else { // need an argument
if (getopt.place >= 0 && getopt.place < args[getopt.ind].length)
getopt.arg = args[getopt.ind].substr(getopt.place);
else if (args.length <= ++getopt.ind) { // no arg
getopt.place = -1;
if (ostr.length > 0 && ostr.charAt(0) == ':') return ':';
return '?';
} else getopt.arg = args[getopt.ind]; // white space
getopt.place = -1;
++getopt.ind;
}
return optopt;
}
/***********************
* Interval operations *
***********************/
Interval = {};
Interval.sort = function(a)
{
if (typeof a[0] == 'number')
a.sort(function(x, y) { return x - y });
else a.sort(function(x, y) { return x[0] != y[0]? x[0] - y[0] : x[1] - y[1] });
}
Interval.merge = function(a, sorted)
{
if (typeof sorted == 'undefined') sorted = true;
if (!sorted) Interval.sort(a);
var k = 0;
for (var i = 1; i < a.length; ++i) {
if (a[k][1] >= a[i][0])
a[k][1] = a[k][1] > a[i][1]? a[k][1] : a[i][1];
else a[++k] = a[i].slice(0);
}
a.length = k + 1;
}
Interval.index_end = function(a, sorted)
{
if (a.length == 0) return;
if (typeof sorted == 'undefined') sorted = true;
if (!sorted) Interval.sort(a);
a[0].push(0);
var k = 0, k_en = a[0][1];
for (var i = 1; i < a.length; ++i) {
if (k_en <= a[i][0]) {
for (++k; k < i; ++k)
if (a[k][1] > a[i][0])
break;
k_en = a[k][1];
}
a[i].push(k);
}
}
Interval.find_intv = function(a, x)
{
var left = -1, right = a.length;
if (typeof a[0] == 'number') {
while (right - left > 1) {
var mid = left + ((right - left) >> 1);
if (a[mid] > x) right = mid;
else if (a[mid] < x) left = mid;
else return mid;
}
} else {
while (right - left > 1) {
var mid = left + ((right - left) >> 1);
if (a[mid][0] > x) right = mid;
else if (a[mid][0] < x) left = mid;
else return mid;
}
}
return left;
}
Interval.find_ovlp = function(a, st, en)
{
if (a.length == 0 || st >= en) return [];
var l = Interval.find_intv(a, st);
var k = l < 0? 0 : a[l][a[l].length - 1];
var b = [];
for (var i = k; i < a.length; ++i) {
if (a[i][0] >= en) break;
else if (st < a[i][1])
b.push(a[i]);
}
return b;
}
/**********************************
* Reverse and reverse complement *
**********************************/
function fasta_read(fn)
{
var h = {}, gt = '>'.charCodeAt(0);
var file = fn == '-'? new File() : new File(fn);
var buf = new Bytes(), seq = null, name = null, seqlen = [];
while (file.readline(buf) >= 0) {
if (buf[0] == gt) {
if (seq != null && name != null) {
seqlen.push([name, seq.length]);
h[name] = seq;
name = seq = null;
}
var m, line = buf.toString();
if ((m = /^>(\S+)/.exec(line)) != null) {
name = m[1];
seq = new Bytes();
}
} else seq.set(buf);
}
if (seq != null && name != null) {
seqlen.push([name, seq.length]);
h[name] = seq;
}
buf.destroy();
file.close();
return [h, seqlen];
}
function fasta_free(fa)
{
for (var name in fa)
fa[name].destroy();
}
Bytes.prototype.reverse = function()
{
for (var i = 0; i < this.length>>1; ++i) {
var tmp = this[i];
this[i] = this[this.length - i - 1];
this[this.length - i - 1] = tmp;
}
}
// reverse complement a DNA string
Bytes.prototype.revcomp = function()
{
if (Bytes.rctab == null) {
var s1 = 'WSATUGCYRKMBDHVNwsatugcyrkmbdhvn';
var s2 = 'WSTAACGRYMKVHDBNwstaacgrymkvhdbn';
Bytes.rctab = [];
for (var i = 0; i < 256; ++i) Bytes.rctab[i] = 0;
for (var i = 0; i < s1.length; ++i)
Bytes.rctab[s1.charCodeAt(i)] = s2.charCodeAt(i);
}
for (var i = 0; i < this.length>>1; ++i) {
var tmp = this[this.length - i - 1];
this[this.length - i - 1] = Bytes.rctab[this[i]];
this[i] = Bytes.rctab[tmp];
}
if (this.length&1)
this[this.length>>1] = Bytes.rctab[this[this.length>>1]];
}
/********************
***** paftools *****
********************/
/*****************
* Miscellaneous *
*****************/
// liftover
function paf_liftover(args)
{
function read_bed(fn, to_merge)
{
if (fn == null) return null;
if (typeof to_merge == 'undefined') to_merge = true;
var file = fn == '-'? new File() : new File(fn);
var buf = new Bytes();
var bed = {};
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
if (bed[t[0]] == null) bed[t[0]] = [];
bed[t[0]].push([parseInt(t[1]), parseInt(t[2])]);
}
buf.destroy();
file.close();
for (var chr in bed) {
Interval.sort(bed[chr]);
if (to_merge)
Interval.merge(bed[chr], true);
Interval.index_end(bed[chr], true);
}
return bed;
}
var re_cigar = /(\d+)([MID])/g, re_tag = /^(\S\S):([AZif]):(\S+)$/;
var c, to_merge = false, min_mapq = 5, min_len = 50000, max_div = 2.0;
var re = /(\d+)([MID])/g;
while ((c = getopt(args, "mq:l:d:")) != null) {
if (c == 'm') to_merge = true;
else if (c == 'q') min_mapq = parseInt(getopt.arg);
else if (c == 'l') min_len = parseInt(getopt.arg);
else if (c == 'd') max_div = parseFloat(getopt.arg);
}
if (args.length - getopt.ind < 2) {
print("Usage: paftools.js liftover [options] <aln.paf> <query.bed>");
print("Options:");
print(" -q INT min mapping quality [" + min_mapq + "]");
print(" -l INT min alignment length [" + min_len + "]");
print(" -d FLOAT max sequence divergence (>=1 to disable) [1]");
exit(1);
}
var bed = read_bed(args[getopt.ind+1], to_merge);
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var buf = new Bytes();
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
if (bed[t[0]] == null) continue; // sequence not present in BED; skip
// parse tp and cg tags
var m, tp = null, cg = null;
for (var i = 12; i < t.length; ++i) {
if ((m = re_tag.exec(t[i])) != null) {
if (m[1] == 'tp') tp = m[3];
else if (m[1] == 'cg') cg = m[3];
}
}
if (tp != 'P' && tp != 'I') continue; // only process primary alignments
if (cg == null) throw Error("unable to find the 'cg' tag");
// filter out bad alignments and check overlaps
for (var i = 1; i <= 3; ++i)
t[i] = parseInt(t[i]);
for (var i = 6; i <= 11; ++i)
t[i] = parseInt(t[i]);
if (t[11] < min_mapq || t[10] < min_len) continue;
var regs = Interval.find_ovlp(bed[t[0]], t[2], t[3]);
if (regs.length == 0) continue; // not overlapping any regions in input BED
if (max_div >= 0.0 && max_div < 1.0) {
var n_gaps = 0, n_opens = 0;
while ((m = re_cigar.exec(cg)) != null)
if (m[2] == 'I' || m[2] == 'D')
n_gaps += parseInt(m[1]), ++n_opens;
var n_mm = t[10] - t[9] - n_gaps;
var n_diff2 = n_mm + n_opens;
if (n_diff2 / (n_diff2 + t[9]) > max_div)
continue;
}
// extract start and end positions
var a = [], r = [], strand = t[4];
for (var i = 0; i < regs.length; ++i) {
var s = regs[i][0], e = regs[i][1];
if (strand == '+') {
a.push([s, 0, i, -2]);
a.push([e - 1, 1, i, -2]);
} else {
a.push([t[1] - e, 0, i, -2]);
a.push([t[1] - s - 1, 1, i, -2]);
}
r.push([-2, -2]);
}
a.sort(function(x, y) { return x[0] - y[0] });
// lift start/end positions
var k = 0, x = t[7], y = strand == '+'? t[2] : t[1] - t[3];
while ((m = re_cigar.exec(cg)) != null) { // TODO: be more careful about edge cases
var len = parseInt(m[1]);
if (m[2] == 'D') { // do nothing for D
x += len;
continue;
}
while (k < a.length && a[k][0] < y) ++k; // skip out-of-range positions
for (var i = k; i < a.length; ++i) {
if (y <= a[i][0] && a[i][0] < y + len)
a[i][3] = m[2] == 'M'? x + (a[i][0] - y) : x;
else break;
}
y += len;
if (m[2] == 'M') x += len;
}
if (x != t[8] || (strand == '+' && y != t[3]) || (strand == '-' && y != t[1] - t[2]))
throw Error("CIGAR is inconsistent with mapping coordinates");
// generate result
for (var i = 0; i < a.length; ++i) {
if (a[i][1] == 0) r[a[i][2]][0] = a[i][3];
else r[a[i][2]][1] = a[i][3] + 1; // change to half-close-half-open
}
for (var i = 0; i < r.length; ++i) {
var name = [t[0], regs[i][0], regs[i][1]].join("_");
if (r[i][0] < 0) name += "_t5", r[i][0] = t[7];
if (r[i][1] < 0) name += "_t3", r[i][1] = t[8];
print(t[5], r[i][0], r[i][1], name, 0, strand);
}
}
buf.destroy();
file.close();
}
// variant calling
function paf_call(args)
{
var re_cs = /([:=*+-])(\d+|[A-Za-z]+)/g, re_tag = /\t(\S\S:[AZif]):(\S+)/g;
var c, min_cov_len = 10000, min_var_len = 50000, gap_thres = 50, gap_thres_long = 1000, min_mapq = 5;
var fa_tmp = null, fa, fa_lens, is_vcf = false, sample_name = "sample";
while ((c = getopt(args, "l:L:g:q:B:f:s:")) != null) {
if (c == 'l') min_cov_len = parseInt(getopt.arg);
else if (c == 'L') min_var_len = parseInt(getopt.arg);
else if (c == 'g') gap_thres = parseInt(getopt.arg);
else if (c == 'G') gap_thres_long = parseInt(getopt.arg);
else if (c == 'q') min_mapq = parseInt(getopt.arg);
else if (c == 'f') fa_tmp = fasta_read(getopt.arg, fa_lens);
else if (c == 's') sample_name = getopt.arg;
}
if (fa_tmp != null) fa = fa_tmp[0], fa_lens = fa_tmp[1], is_vcf = true;
if (args.length == getopt.ind) {
print("Usage: sort -k6,6 -k8,8n <with-cs.paf> | paftools.js call [options] -");
print("Options:");
print(" -l INT min alignment length to compute coverage ["+min_cov_len+"]");
print(" -L INT min alignment length to call variants ["+min_var_len+"]");
print(" -q INT min mapping quality ["+min_mapq+"]");
print(" -g INT short/long gap threshold (for statistics only) ["+gap_thres+"]");
print(" -f FILE reference sequences (enabling VCF output) [null]");
print(" -s NAME sample name in VCF header ["+sample_name+"]");
exit(1);
}
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var buf = new Bytes();
var tot_len = 0, n_sub = [0, 0, 0], n_ins = [0, 0, 0, 0, 0], n_del = [0, 0, 0, 0, 0];
function print_vcf(o, fa)
{
var v = null;
if (o[3] != 1) return; // coverage is one; skip
if (o[5] == '-' && o[6] == '-') return;
if (o[5] != '-' && o[6] != '-') { // snp
v = [o[0], o[1] + 1, '.', o[5].toUpperCase(), o[6].toUpperCase()];
} else if (o[1] > 0) { // shouldn't happen in theory
if (fa[o[0]] == null) throw Error('sequence "' + o[0] + '" is absent from the reference FASTA');
if (o[1] >= fa[o[0]].length) throw Error('position ' + o[1] + ' exceeds the length of sequence "' + o[0] + '"');
var ref = String.fromCharCode(fa[o[0]][o[1]-1]).toUpperCase();
if (o[5] == '-') // insertion
v = [o[0], o[1], '.', ref, ref + o[6].toUpperCase()];
else // deletion
v = [o[0], o[1], '.', ref + o[5].toUpperCase(), ref];
}
v.push(o[4], '.', 'QNAME=' + o[7] + ';QSTART=' + (o[8]+1) + ';QSTRAND=' + (rev? '-' : '+'), 'GT', '1/1');
if (v == null) throw Error("unexpected variant: [" + o.join(",") + "]");
print(v.join("\t"));
}
function count_var(o)
{
if (o[3] > 1) return;
if (o[5] == '-' && o[6] == '-') return;
if (o[5] == '-') { // insertion
var l = o[6].length;
if (l == 1) ++n_ins[0];
else if (l == 2) ++n_ins[1];
else if (l < gap_thres) ++n_ins[2];
else if (l < gap_thres_long) ++n_ins[3];
else ++n_ins[4];
} else if (o[6] == '-') { // deletion
var l = o[5].length;
if (l == 1) ++n_del[0];
else if (l == 2) ++n_del[1];
else if (l < gap_thres) ++n_del[2];
else if (l < gap_thres_long) ++n_del[3];
else ++n_del[4];
} else {
++n_sub[0];
var s = (o[5] + o[6]).toLowerCase();
if (s == 'ag' || s == 'ga' || s == 'ct' || s == 'tc')
++n_sub[1];
else ++n_sub[2];
}
}
if (is_vcf) {
print('##fileformat=VCFv4.1');
for (var i = 0; i < fa_lens.length; ++i)
print('##contig=<ID=' + fa_lens[i][0] + ',length=' + fa_lens[i][1] + '>');
print('##INFO=<ID=QNAME,Number=1,Type=String,Description="Query name">');
print('##INFO=<ID=QSTART,Number=1,Type=Integer,Description="Query start">');
print('##INFO=<ID=QSTRAND,Number=1,Type=String,Description="Query strand">');
print('##FORMAT=<ID=GT,Number=1,Type=String,Description="Genotype">');
print('#CHROM POS ID REF ALT QUAL FILTER INFO FORMAT '+sample_name);
}
var a = [], out = [];
var c1_ctg = null, c1_start = 0, c1_end = 0, c1_counted = false, c1_len = 0;
while (file.readline(buf) >= 0) {
var line = buf.toString();
var m, t = line.split("\t", 12);
if (t.length < 12 || t[5] == '*') continue; // unmapped
for (var i = 6; i <= 11; ++i)
t[i] = parseInt(t[i]);
if (t[10] < min_cov_len || t[11] < min_mapq) continue;
//print(t[0], t[7], t[8], c1_start, c1_end);
for (var i = 1; i <= 3; ++i)
t[i] = parseInt(t[i]);
var ctg = t[5], x = t[7], end = t[8];
var query = t[0], rev = (t[4] == '-'), y = rev? t[3] : t[2];
// collect tags
var cs = null, tp = null, have_s1 = false, have_s2 = false;
while ((m = re_tag.exec(line)) != null) {
if (m[1] == 'cs:Z') cs = m[2];
else if (m[1] == 'tp:A') tp = m[2];
else if (m[1] == 's1:i') have_s1 = true;
else if (m[1] == 's2:i') have_s2 = true;
}
if (have_s1 && !have_s2) continue;
if (tp != null && (tp == 'S' || tp == 'i')) continue;
// compute regions covered by 1 contig
if (ctg != c1_ctg || x >= c1_end) {
if (c1_counted && c1_end > c1_start) {
c1_len += c1_end - c1_start;
if (!is_vcf) print('R', c1_ctg, c1_start, c1_end);
}
c1_ctg = ctg, c1_start = x, c1_end = end;
c1_counted = (t[10] >= min_var_len);
} else if (end > c1_end) { // overlap
if (c1_counted && x > c1_start) {
c1_len += x - c1_start;
if (!is_vcf) print('R', c1_ctg, c1_start, x);
}
c1_start = c1_end, c1_end = end;
c1_counted = (t[10] >= min_var_len);
} else if (end > c1_start) { // contained
if (c1_counted && x > c1_start) {
c1_len += x - c1_start;
if (!is_vcf) print('R', c1_ctg, c1_start, x);
}
c1_start = end;
} // else, the alignment precedes the cov1 region; do nothing
// output variants ahead of this alignment
while (out.length) {
if (out[0][0] != ctg || out[0][2] <= x) {
count_var(out[0]);
if (is_vcf) print_vcf(out[0], fa);
else print('V', out[0].join("\t"));
out.shift();
} else break;
}
// update coverage
for (var i = 0; i < out.length; ++i)
if (out[i][1] >= x && out[i][2] <= end)
++out[i][3];
// drop alignments that don't overlap with the current one
var k = 0;
for (var i = 0; i < a.length; ++i)
if (a[i][0] == ctg && a[i][2] > x)
a[k++] = a[i];
a.length = k;
// core loop
if (t[10] >= min_var_len) {
if (cs == null) continue; // no cs tag
var blen = 0, n_diff = 0;
tot_len += t[10];
while ((m = re_cs.exec(cs)) != null) {
var cov = 1;
if (m[1] == '*' || m[1] == '+' || m[1] == '-')
for (var i = 0; i < a.length; ++i)
if (a[i][2] > x) ++cov;
var qs, qe;
if (m[1] == '=' || m[1] == ':') {
var l = m[1] == '='? m[2].length : parseInt(m[2]);
if (rev) y -= l;
else y += l;
x += l, blen += l;
} else if (m[1] == '*') {
if (rev) qs = y - 1, qe = y, --y;
else qs = y, qe = y + 1, ++y;
var br = m[2].charAt(0), bq = m[2].charAt(1);
if (br != 'n' && bq != 'n') { // don't call a SNP if there is an ambiguous base
out.push([t[5], x, x+1, cov, t[11], br, bq, query, qs, qe, rev? '-' : '+']);
++n_diff;
}
++x, ++blen;
} else if (m[1] == '+') {
var l = m[2].length;
if (rev) qs = y - l, qe = y, y -= l;
else qs = y, qe = y + l, y += l;
out.push([t[5], x, x, cov, t[11], '-', m[2], query, qs, qe, rev? '-' : '+']);
++blen, ++n_diff;
} else if (m[1] == '-') {
var l = m[2].length;
out.push([t[5], x, x + l, cov, t[11], m[2], '-', query, y, y, rev? '-' : '+']);
x += l, ++blen, ++n_diff;
}
}
}
a.push([t[5], t[7], t[8]]);
}
if (c1_counted && c1_end > c1_start) {
c1_len += c1_end - c1_start;
if (!is_vcf) print('R', c1_ctg, c1_start, c1_end);
}
while (out.length) {
count_var(out[0]);
if (is_vcf) print_vcf(out[0], fa);
else print('V', out[0].join("\t"));
out.shift();
}
//warn(tot_len + " alignment columns considered in calling");
warn(c1_len + " reference bases covered by exactly one contig");
warn(n_sub[0] + " substitutions; ts/tv = " + (n_sub[1]/n_sub[2]).toFixed(3));
warn(n_del[0] + " 1bp deletions");
warn(n_ins[0] + " 1bp insertions");
warn(n_del[1] + " 2bp deletions");
warn(n_ins[1] + " 2bp insertions");
warn(n_del[2] + " [3,"+gap_thres+") deletions");
warn(n_ins[2] + " [3,"+gap_thres+") insertions");
warn(n_del[3] + " ["+gap_thres+","+gap_thres_long+") deletions");
warn(n_ins[3] + " ["+gap_thres+","+gap_thres_long+") insertions");
warn(n_del[4] + " >=" + gap_thres_long + " deletions");
warn(n_ins[4] + " >=" + gap_thres_long + " insertions");
buf.destroy();
file.close();
if (fa != null) fasta_free(fa);
}
function paf_asmstat(args)
{
var c, min_query_len = 0, min_seg_len = 10000, max_diff = 0.01, bp_flank_len = 0, bp_gap_len = 0;
while ((c = getopt(args, "l:d:b:g:q:")) != null) {
if (c == 'l') min_seg_len = parseInt(getopt.arg);
else if (c == 'd') max_diff = parseFloat(getopt.arg);
else if (c == 'b') bp_flank_len = parseInt(getopt.arg);
else if (c == 'g') bp_gap_len = parseInt(getopt.arg);
else if (c == 'q') min_query_len = parseInt(getopt.arg);
}
if (getopt.ind == args.length) {
print("Usage: paftools.js asmstat [options] <ref.fa.fai> <asm1.paf> [...]");
print("Options:");
print(" -q INT ignore query shorter than INT [0]");
print(" -l INT min alignment block length [" + min_seg_len + "]");
print(" -d FLOAT max gap-compressed sequence divergence [" + max_diff + "]");
exit(1);
}
var file, buf = new Bytes();
var ref_len = 0;
file = new File(args[getopt.ind]);
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
ref_len += parseInt(t[1]);
}
file.close();
function process_query(qblocks, qblock_len, bp, qi) {
qblocks.sort(function(a,b) { return a[0]-b[0]; });
var last_k = null, last_blen = null, st = -1, en = -1, qcov = 0;
for (var k = 0; k < qblocks.length; ++k) {
var blen = qblocks[k][1] - qblocks[k][0];
if (k > 0 && qblocks[k][0] < qblocks[k-1][1]) {
if (qblocks[k][1] < qblocks[k-1][1]) continue;
blen = qblocks[k][1] - qblocks[k-1][1];
}
qblock_len.push(blen);
if (qblocks[k][0] > en) {
qcov += en - st;
st = qblocks[k][0];
en = qblocks[k][1];
} else en = en > qblocks[k][1]? en : qblocks[k][1];
if (last_k != null) {
var gap = 1000000000;
if (qblocks[k][2] == qblocks[last_k][2] && qblocks[k][3] == qblocks[last_k][3]) { // same chr and strand
var g1 = qblocks[k][0] - qblocks[last_k][1];
var g2 = qblocks[k][2] == '+'? qblocks[k][4] - qblocks[last_k][5] : qblocks[last_k][4] - qblocks[k][5];
gap = g1 > g2? g1 - g2 : g2 - g1;
}
var min = blen < last_blen? blen : last_blen;
var flank = k == 0? min : blen;
bp.push([flank, gap]);
qi.bp.push([flank, gap]);
}
last_k = k, last_blen = blen;
}
qcov += en - st;
return qcov;
}
function N50(lens, tot, quantile) {
lens.sort(function(a,b) { return b - a; });
if (tot == null) {
tot = 0;
for (var k = 0; k < lens.length; ++k)
tot += lens[k];
}
var sum = 0;
for (var k = 0; k < lens.length; ++k) {
if (sum <= quantile * tot && sum + lens[k] > quantile * tot)
return lens[k];
sum += lens[k];
}
}
function count_bp(bp, min_blen, min_gap) {
var n_bp = 0;
for (var k = 0; k < bp.length; ++k)
if (bp[k][0] >= min_blen && bp[k][1] >= min_gap)
++n_bp;
return n_bp;
}
function compute_diff(cigar, NM) {
var m, re = /(\d+)([MID])/g;
var n_M = 0, n_gapo = 0, n_gaps = 0;
while ((m = re.exec(cigar)) != null) {
var len = parseInt(m[1]);
if (m[2] == 'M') n_M += len;
else ++n_gapo, n_gaps += len;
}
if (NM < n_gaps) throw Error('NM is smaller the number of gaps');
return (NM - n_gaps + n_gapo) / (n_M + n_gapo);
}
var labels = ['Length', 'l_cov', 'Rcov', 'Rdup', 'Qcov', 'NG75', 'NG50', 'NGA50', '#breaks', 'bp(' + min_seg_len + ',0)', 'bp(' + min_seg_len + ',10k)'];
var rst = [];
for (var i = 0; i < labels.length; ++i)
rst[i] = [];
var n_asm = args.length - (getopt.ind + 1);
var header = ["Metric"];
for (var i = 0; i < n_asm; ++i) {
var n_breaks = 0, qcov = 0;
var fn = args[getopt.ind + 1 + i];
var label = fn.replace(/.paf(.gz)?$/, "");
header.push(label);
var ref_blocks = [], qblock_len = [], qblocks = [], bp = [];
var query = {}, qinfo = {};
var last_qname = null;
file = new File(fn);
while (file.readline(buf) >= 0) {
var m, line = buf.toString();
var t = line.split("\t");
t[1] = parseInt(t[1]);
if (t[1] < min_query_len) continue;
if (t.length < 2) continue;
query[t[0]] = t[1];
if (qinfo[t[0]] == null) qinfo[t[0]] = {};
qinfo[t[0]].len = t[1];
qinfo[t[0]].bp = [];
if (t.length < 9 || t[5] == "*") continue;
if (!/\ttp:A:[PI]/.test(line)) continue;
if ((m = /\tcg:Z:(\S+)/.exec(line)) == null) continue;
var cigar = m[1];
if ((m = /\tNM:i:(\d+)/.exec(line)) == null) continue;
var NM = parseInt(m[1]);
var diff = compute_diff(cigar, NM);
t[2] = parseInt(t[2]);
t[3] = parseInt(t[3]);
t[7] = parseInt(t[7]);
t[8] = parseInt(t[8]);
if (t[0] == last_qname) ++n_breaks;
if (diff > max_diff) continue;
if (t[3] - t[2] < min_seg_len) continue;
if (t[0] != last_qname) {
if (last_qname != null)
qcov += process_query(qblocks, qblock_len, bp, qinfo[last_qname]);
qblocks = [];
last_qname = t[0];
}
ref_blocks.push([t[5], t[7], t[8]]);
qblocks.push([t[2], t[3], t[4], t[5], t[7], t[8]]);
}
if (last_qname != null)
qcov += process_query(qblocks, qblock_len, bp, qinfo[last_qname]);
file.close();
// compute NG50
var asm_len = 0, asm_lens = []
for (var ctg in query) {
asm_len += query[ctg];
asm_lens.push(query[ctg]);
}
rst[0][i] = asm_len;
rst[5][i] = N50(asm_lens, ref_len, 0.75);
rst[6][i] = N50(asm_lens, ref_len, 0.5);
// compute coverage
var l_cov = 0;
ref_blocks.sort(function(a, b) { return a[0] > b[0]? 1 : a[0] < b[0]? -1 : a[1] - b[1]; });
var last_ref = null, st = -1, en = -1;
for (var j = 0; j < ref_blocks.length; ++j) {
if (ref_blocks[j][0] != last_ref || ref_blocks[j][1] > en) {
l_cov += en - st;
last_ref = ref_blocks[j][0];
st = ref_blocks[j][1];
en = ref_blocks[j][2];
} else en = en > ref_blocks[j][2]? en : ref_blocks[j][2];
}
l_cov += en - st;
rst[1][i] = l_cov;
rst[2][i] = (100.0 * (l_cov / ref_len)).toFixed(2) + '%';
rst[4][i] = (100.0 * (qcov / asm_len)).toFixed(2) + '%';
// compute cov1 and cov2+ lengths; see paf_call() for details
var c1_ctg = null, c1_start = 0, c1_end = 0, c1_len = 0;
for (var j = 0; j < ref_blocks.length; ++j) {
if (ref_blocks[j][0] != c1_ctg || ref_blocks[j][1] >= c1_end) {
if (c1_end > c1_start)
c1_len += c1_end - c1_start;
c1_ctg = ref_blocks[j][0], c1_start = ref_blocks[j][1], c1_end = ref_blocks[j][2];
} else if (ref_blocks[j][2] > c1_end) { // overlap
if (ref_blocks[j][1] > c1_start)
c1_len += ref_blocks[j][1] - c1_start;
c1_start = c1_end, c1_end = ref_blocks[j][2];
} else if (ref_blocks[j][2] > c1_start) { // contained
if (ref_blocks[j][1] > c1_start)
c1_len += ref_blocks[j][1] - c1_start;
c1_start = ref_blocks[j][2];
}
//print(ref_blocks[j][0], ref_blocks[j][1], ref_blocks[j][2], c1_start, c1_end, c1_len);
}
if (c1_end > c1_start)
c1_len += c1_end - c1_start;
rst[3][i] = (100 * (l_cov - c1_len) / l_cov).toFixed(2) + '%';
// compute NGA50
rst[7][i] = N50(qblock_len, ref_len, 0.5);
// compute break points
rst[8][i] = n_breaks;
rst[9][i] = count_bp(bp, 500, 0);
rst[10][i] = count_bp(bp, 500, 10000);
// nb-plot; NOT USED
/*
var qa = [];
for (var qn in qinfo)
qa.push([qinfo[qn].len, qinfo[qn].bp]);
qa = qa.sort(function(a, b) { return b[0] - a[0] });
var sum = 0, n_bp = 0, next_quantile = 0.1;
for (var j = 0; j < qa.length; ++j) {
sum += qa[j][0];
for (var k = 0; k < qa[j][1].length; ++k)
if (qa[j][1][k][0] >= bp_flank_len && qa[j][1][k][1] >= bp_gap_len)
++n_bp;
if (sum >= ref_len * next_quantile) {
print(label, Math.floor(next_quantile * 100 + .5), qa[j][0], (sum / n_bp).toFixed(0), n_bp);
next_quantile += 0.1;
if (next_quantile >= 1.0) break;
}
}
*/
}
buf.destroy();
if (bp_flank_len <= 0) {
print(header.join("\t"));
for (var i = 0; i < labels.length; ++i)
print(labels[i], rst[i].join("\t"));
}
}
function paf_asmgene(args)
{
var c, opt = { min_cov:0.99, min_iden:0.99 }, print_err = false, auto_only = false;
while ((c = getopt(args, "i:c:ea")) != null)
if (c == 'i') opt.min_iden = parseFloat(getopt.arg);
else if (c == 'c') opt.min_cov = parseFloat(getopt.arg);
else if (c == 'e') print_err = true;
else if (c == 'a') auto_only = true;
var n_fn = args.length - getopt.ind;
if (n_fn < 2) {
print("Usage: paftools.js asmgene [options] <ref-splice.paf> <asm-splice.paf> [...]");
print("Options:");
print(" -i FLOAT min identity [" + opt.min_iden + "]");
print(" -c FLOAT min coverage [" + opt.min_cov + "]");
print(" -a only evaluate genes mapped to the autosomes");
print(" -e print fragmented/missing genes");
exit(1);
}
function process_query(opt, a) {
var b = [], cnt = [0, 0, 0];
for (var j = 0; j < a.length; ++j) {
if (a[j][4] < a[j][5] * opt.min_iden)
continue;
b.push(a[j].slice(0));
}
if (b.length == 0) return cnt;
// count full
var n_full = 0;
for (var j = 0; j < b.length; ++j)
if (b[j][3] - b[j][2] >= b[j][1] * opt.min_cov)
++n_full;
cnt[0] = n_full;
// compute coverage
b = b.sort(function(x, y) { return x[2] - y[2] });
var l_cov = 0, st = b[0][2], en = b[0][3];
for (var j = 1; j < b.length; ++j) {
if (b[j][2] <= en)
en = b[j][3] > en? b[j][3] : en;
else l_cov += en - st;
}
l_cov += en - st;
cnt[1] = l_cov / b[0][1];
cnt[2] = b.length;
return cnt;
}
var buf = new Bytes();
var gene = {}, header = [], refpos = {};
for (var i = getopt.ind; i < args.length; ++i) {
var fn = args[i];
var label = fn.replace(/.paf(.gz)?$/, "");
header.push(label);
var file = new File(fn), a = [];
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
var ql = parseInt(t[1]), qs = parseInt(t[2]), qe = parseInt(t[3]), mlen = parseInt(t[9]), blen = parseInt(t[10]), mapq = parseInt(t[11]);
if (i == getopt.ind) refpos[t[0]] = [t[0], t[1], t[5], t[7], t[8]];
if (gene[t[0]] == null) gene[t[0]] = [];
if (a.length && t[0] != a[0][0]) {
gene[a[0][0]][i - getopt.ind] = process_query(opt, a);
a = [];
}
a.push([t[0], ql, qs, qe, mlen, blen]);
}
if (a.length)
gene[t[0]][i - getopt.ind] = process_query(opt, a);
file.close();
}
// select the longest genes (not optimal, but should be good enough)
var gene_list = [], gene_nr = {};
for (var g in refpos)
gene_list.push(refpos[g]);
gene_list = gene_list.sort(function(a, b) { return a[2] < b[2]? -1 : a[2] > b[2]? 1 : a[3] - b[3] });
var last = 0;
for (var j = 1; j < gene_list.length; ++j) {
if (gene_list[j][2] != gene_list[last][2] || gene_list[j][3] >= gene_list[last][4]) {
gene_nr[gene_list[last][0]] = 1;
last = j;
} else if (gene_list[j][1] > gene_list[last][1]) {
last = j;
}
}
gene_nr[gene_list[last][0]] = 1;
// count and print
var col1 = ["full_sgl", "full_dup", "frag", "part50+", "part10+", "part10-"];
var rst = [];
for (var k = 0; k < col1.length; ++k) {
rst[k] = [];
for (var i = 0; i < n_fn; ++i)
rst[k][i] = 0;
}
for (var g in gene) {
if (gene[g][0] == null || gene[g][0][0] != 1) continue;
if (gene_nr[g] == null) continue;
if (auto_only && /^(chr)?[XY]$/.test(refpos[g][2])) continue;
for (var i = 0; i < n_fn; ++i) {
if (gene[g][i] == null) {
rst[4][i]++;
if (print_err) print('M', header[i], refpos[g].join("\t"));
} else if (gene[g][i][0] == 1) rst[0][i]++;
else if (gene[g][i][0] > 1) {
rst[1][i]++;
if (print_err) print('D', header[i], refpos[g].join("\t"));
} else if (gene[g][i][1] >= opt.min_cov) {
rst[2][i]++;
if (print_err) print('F', header[i], refpos[g].join("\t"));
} else if (gene[g][i][1] >= 0.5) {
rst[3][i]++;
if (print_err) print('5', header[i], refpos[g].join("\t"));
} else if (gene[g][i][1] >= 0.1) {
rst[4][i]++;
if (print_err) print('1', header[i], refpos[g].join("\t"));
} else {
rst[5][i]++;
if (print_err) print('0', header[i], refpos[g].join("\t")); // TODO: reduce code duplicates...
}
}
}
print('H', 'Metric', header.join("\t"));
for (var k = 0; k < rst.length; ++k) {
print('X', col1[k], rst[k].join("\t"));
}
buf.destroy();
}
function paf_stat(args)
{
var c, gap_out_len = null;
while ((c = getopt(args, "l:")) != null)
if (c == 'l') gap_out_len = parseInt(getopt.arg);
if (getopt.ind == args.length) {
print("Usage: paftools.js stat [-l gapOutLen] <in.sam>|<in.paf>");
exit(1);
}
var buf = new Bytes();
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var re = /(\d+)([MIDSHNX=])/g;
var lineno = 0, n_pri = 0, n_2nd = 0, n_seq = 0, n_cigar_64k = 0, l_tot = 0, l_cov = 0;
var n_gap = [[0, 0, 0, 0, 0, 0], [0, 0, 0, 0, 0, 0]];
function cov_len(regs)
{
regs.sort(function(a,b) {return a[0]-b[0]});
var st = regs[0][0], en = regs[0][1], l = 0;
for (var i = 1; i < regs.length; ++i) {
if (regs[i][0] < en)
en = en > regs[i][1]? en : regs[i][1];
else l += en - st, st = regs[i][0], en = regs[i][1];
}
l += en - st;
return l;
}
var last = null, last_qlen = null, regs = [];
while (file.readline(buf) >= 0) {
var line = buf.toString();
++lineno;
if (line.charAt(0) != '@') {
var t = line.split("\t", 12);
var m, rs, cigar = null, is_pri = false, is_sam = false, is_rev = false, tname = null;
var atlen = null, aqlen, qs, qe, mapq, ori_qlen;
if (t.length < 2) continue;
if (t[4] == '+' || t[4] == '-' || t[4] == '*') { // PAF
if (t[4] == '*') continue; // unmapped
if (!/\ts2:i:\d+/.test(line)) {
++n_2nd;
continue;
}
if ((m = /\tcg:Z:(\S+)/.exec(line)) != null)
cigar = m[1];
if (cigar == null) {
warn("WARNING: no CIGAR at line " + lineno);
continue;
}
tname = t[5];
qs = parseInt(t[2]), qe = parseInt(t[3]);
aqlen = qe - qs;
is_rev = t[4] == '+'? false : true;
rs = parseInt(t[7]);
atlen = parseInt(t[8]) - rs;
mapq = parseInt(t[11]);
ori_qlen = parseInt(t[1]);
} else { // SAM
var flag = parseInt(t[1]);
if ((flag & 4) || t[2] == '*' || t[5] == '*') continue;
if (flag & 0x100) {
++n_2nd;
continue;
}
cigar = t[5];
tname = t[2];
rs = parseInt(t[3]) - 1;
mapq = parseInt(t[4]);
aqlen = t[9].length;
is_sam = true;
is_rev = !!(flag&0x10);
}
++n_pri;
if (last != t[0]) {
if (last != null) {
l_tot += last_qlen;
l_cov += cov_len(regs);
}
regs = [];
++n_seq, last = t[0];
}
var M = 0, tl = 0, ql = 0, clip = [0, 0], n_cigar = 0, sclip = 0;
while ((m = re.exec(cigar)) != null) {
var l = parseInt(m[1]);
++n_cigar;
if (m[2] == 'M' || m[2] == '=' || m[2] == 'X') {
tl += l, ql += l, M += l;
} else if (m[2] == 'I' || m[2] == 'D') {
var type;
if (l < 50) type = 0;
else if (l < 100) type = 1;
else if (l < 300) type = 2;
else if (l < 400) type = 3;
else if (l < 1000) type = 4;
else type = 5;
if (m[2] == 'I') ql += l, ++n_gap[0][type];
else tl += l, ++n_gap[1][type];
if (gap_out_len != null && l >= gap_out_len)
print(t[0], ql, is_rev? '-' : '+', tname, rs + tl, m[2], l);
} else if (m[2] == 'N') {
tl += l;
} else if (m[2] == 'S') {
clip[M == 0? 0 : 1] = l, sclip += l;
} else if (m[2] == 'H') {
clip[M == 0? 0 : 1] = l;
}
}
if (n_cigar > 65535) ++n_cigar_64k;
if (ql + sclip != aqlen)
warn("WARNING: aligned query length is inconsistent with CIGAR at line " + lineno + " (" + (ql+sclip) + " != " + aqlen + ")");
if (atlen != null && atlen != tl)
warn("WARNING: aligned reference length is inconsistent with CIGAR at line " + lineno);
if (is_sam) {
qs = clip[is_rev? 1 : 0], qe = qs + ql;
ori_qlen = clip[0] + ql + clip[1];
}
regs.push([qs, qe]);
last_qlen = ori_qlen;
}
}
if (regs.length) {
l_tot += last_qlen;
l_cov += cov_len(regs);
}
file.close();
buf.destroy();
if (gap_out_len == null) {
print("Number of mapped sequences: " + n_seq);
print("Number of primary alignments: " + n_pri);
print("Number of secondary alignments: " + n_2nd);
print("Number of primary alignments with >65535 CIGAR operations: " + n_cigar_64k);
print("Number of bases in mapped sequences: " + l_tot);
print("Number of mapped bases: " + l_cov);
print("Number of insertions in [0,50): " + n_gap[0][0]);
print("Number of insertions in [50,100): " + n_gap[0][1]);
print("Number of insertions in [100,300): " + n_gap[0][2]);
print("Number of insertions in [300,400): " + n_gap[0][3]);
print("Number of insertions in [400,1000): " + n_gap[0][4]);
print("Number of insertions in [1000,inf): " + n_gap[0][5]);
print("Number of deletions in [0,50): " + n_gap[1][0]);
print("Number of deletions in [50,100): " + n_gap[1][1]);
print("Number of deletions in [100,300): " + n_gap[1][2]);
print("Number of deletions in [300,400): " + n_gap[1][3]);
print("Number of deletions in [400,1000): " + n_gap[1][4]);
print("Number of deletions in [1000,inf): " + n_gap[1][5]);
}
}
function paf_bedcov(args)
{
function read_bed(fn, to_merge, to_dedup)
{
var file = new File(fn);
var buf = new Bytes();
var h = {};
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
if (h[t[0]] == null)
h[t[0]] = [];
var bst = parseInt(t[1]);
var ben = parseInt(t[2]);
if (t.length >= 12 && /^\d+$/.test(t[9])) {
t[9] = parseInt(t[9]);
var sz = t[10].split(",");
var st = t[11].split(",");
for (var i = 0; i < t[9]; ++i) {
st[i] = parseInt(st[i]);
sz[i] = parseInt(sz[i]);
h[t[0]].push([bst + st[i], bst + st[i] + sz[i], 0, 0, 0]);
}
} else {
h[t[0]].push([bst, ben, 0, 0, 0]);
}
}
buf.destroy();
file.close();
for (var chr in h) {
if (to_merge) Interval.merge(h[chr], false);
else if (to_dedup) Interval.dedup(h[chr], false);
else Interval.sort(h[chr]);
Interval.index_end(h[chr]);
}
return h;
}
var c, print_len = false, to_merge = true, to_dedup = false, fn_excl = null;
while ((c = getopt(args, "pde:")) != null) {
if (c == 'p') print_len = true;
else if (c == 'd') to_dedup = true, to_merge = false;
else if (c == 'e') fn_excl = getopt.arg;
}
if (args.length - getopt.ind < 2) {
print("Usage: paftools.js bedcov [options] <regions.bed> <target.bed>");
print("Options:");
print(" -e FILE exclude target regions (2nd file) overlapping BED FILE []");
print(" -p print number of covered bases for each target");
exit(1);
}
var excl = fn_excl != null? read_bed(fn_excl, true, false) : null;
var target = read_bed(args[getopt.ind], to_merge, to_dedup);
var file, buf = new Bytes();
var tot_len = 0, hit_len = 0;
file = args[getopt.ind+1] != '-'? new File(args[getopt.ind+1]) : new File();
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
var a = [];
var bst = parseInt(t[1]);
var ben = parseInt(t[2]);
if (t.length >= 12 && /^\d+$/.test(t[9])) { // BED12
t[9] = parseInt(t[9]);
var sz = t[10].split(",");
var st = t[11].split(",");
for (var i = 0; i < t[9]; ++i) {
st[i] = parseInt(st[i]);
sz[i] = parseInt(sz[i]);
a.push([bst + st[i], bst + st[i] + sz[i], false]);
}
} else a.push([bst, ben, false]); // 3-column BED
var feat_len = 0;
for (var i = 0; i < a.length; ++i) {
if (excl != null && excl[t[0]] != null) {
var oe = Interval.find_ovlp(excl[t[0]], a[i][0], a[i][1]);
if (oe.length > 0)
continue;
}
a[i][2] = true;
feat_len += a[i][1] - a[i][0];
}
tot_len += feat_len;
if (target[t[0]] == null) continue;
var b = [];
for (var i = 0; i < a.length; ++i) {
if (!a[i][2]) continue;
var o = Interval.find_ovlp(target[t[0]], a[i][0], a[i][1]);
for (var j = 0; j < o.length; ++j) {
var max_st = o[j][0] > a[i][0]? o[j][0] : a[i][0];
var min_en = o[j][1] < a[i][1]? o[j][1] : a[i][1];
b.push([max_st, min_en]);
o[j][2] += min_en - max_st;
++o[j][3];
if (max_st == o[j][0] && min_en == o[j][1])
++o[j][4];
}
}
// find the length covered
var feat_hit_len = 0;
if (b.length > 0) {
b.sort(function(a,b) {return a[0]-b[0]});
var st = b[0][0], en = b[0][1];
for (var i = 1; i < b.length; ++i) {
if (b[i][0] <= en) en = en > b[i][1]? en : b[i][1];
else feat_hit_len += en - st, st = b[i][0], en = b[i][1];
}
feat_hit_len += en - st;
}
hit_len += feat_hit_len;
if (print_len) print('F', t.slice(0, 4).join("\t"), feat_len, feat_hit_len);
}
file.close();
buf.destroy();
warn("# target bases: " + tot_len);
warn("# target bases overlapping regions: " + hit_len + ' (' + (100.0 * hit_len / tot_len).toFixed(2) + '%)');
}
function paf_vcfpair(args)
{
var c, is_male = false, sample = 'syndip', hgver = null;
var PAR = { '37':[[0, 2699520], [154931043, 155260560]] };
while ((c = getopt(args, "ms:g:")) != null) {
if (c == 'm') is_male = true;
else if (c == 's') sample = getopt.arg;
else if (c == 'g') hgver = getopt.arg;
}
if (is_male && (hgver == null || PAR[hgver] == null))
throw("for a male, -g must be specified to properly handle PARs on chrX");
if (getopt.ind == args.length) {
print("Usage: paftools.js vcfpair [options] <in.pair.vcf>");
print("Options:");
print(" -m the sample is male");
print(" -g STR human genome version '37' []");
print(" -s STR sample name [" + sample + "]");
exit(1);
}
var re_ctg = is_male? /^(chr)?([0-9]+|X|Y)$/ : /^(chr)?([0-9]+|X)$/;
var label = ['1', '2'];
var buf = new Bytes();
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
while (file.readline(buf) >= 0) {
var m, line = buf.toString();
if (line.charAt(0) == '#') {
if (/^##(source|reference)=/.test(line)) continue;
if ((m = /^##contig=.*ID=([^\s,]+)/.exec(line)) != null) {
if (!re_ctg.test(m[1])) continue;
} else if (/^#CHROM/.test(line)) {
var t = line.split("\t");
--t.length;
t[t.length-1] = sample;
line = t.join("\t");
print('##FILTER=<ID=HET1,Description="Heterozygous in the first haplotype">');
print('##FILTER=<ID=HET2,Description="Heterozygous in the second haplotype">');
print('##FILTER=<ID=GAP1,Description="Uncalled in the first haplotype">');
print('##FILTER=<ID=GAP2,Description="Uncalled in the second haplotype">');
}
print(line);
continue;
}
var t = line.split("\t");
if (!re_ctg.test(t[0])) continue;
var GT = null, AD = null, FILTER = [], HT = [null, null];
for (var i = 0; i < 2; ++i) {
if ((m = /^(\.|[0-9]+)\/(\.|[0-9]+):(\S+)/.exec(t[9+i])) == null) {
warn(line);
throw Error("malformatted VCF");
}
var s = m[3].split(",");
if (AD == null) {
AD = [];
for (var j = 0; j < s.length; ++j)
AD[j] = 0;
}
for (var j = 0; j < s.length; ++j)
AD[j] += parseInt(s[j]);
if (m[1] == '.') {
FILTER.push('GAP' + label[i]);
HT[i] = '.';
} else if (m[1] != m[2]) {
FILTER.push('HET' + label[i]);
HT[i] = '.';
} else HT[i] = m[1];
}
--t.length;
// test if this is in a haploid region
var hap = 0, st = parseInt(t[1]), en = st + t[3].length;
if (is_male) {
if (/^(chr)?X/.test(t[0])) {
if (hgver != null && PAR[hgver] != null) {
var r = PAR[hgver], in_par = false;
for (var i = 0; i < r.length; ++i)
if (r[i][0] <= st && en <= r[i][1])
in_par = true;
hap = in_par? 0 : 2;
}
} else if (/^(chr)?Y/.test(t[0])) {
hap = 1;
}
}
// special treatment for haploid regions
if (hap > 0 && FILTER.length == 1) {
if ((hap == 2 && FILTER[0] == "GAP1") || (hap == 1 && FILTER[0] == "GAP2"))
FILTER.length = 0;
}
// update VCF
t[5] = 30; // fake QUAL
t[6] = FILTER.length? FILTER.join(";") : ".";
t[9] = HT.join("|") + ":" + AD.join(",");
print(t.join("\t"));
}
file.close();
buf.destroy();
}
/**********************
* Conversion related *
**********************/
function paf_view(args)
{
var c, line_len = 80, fmt = "aln";
while ((c = getopt(args, "f:l:")) != null) {
if (c == 'f') {
fmt = getopt.arg;
if (fmt != "aln" && fmt != "lastz-cigar" && fmt != "maf")
throw Error("format must be one of aln, lastz-cigar and maf");
} else if (c == 'l') line_len = parseInt(getopt.arg);
}
if (line_len == 0) line_len = 0x7fffffff;
if (getopt.ind == args.length) {
print("Usage: paftools.js view [options] <in.paf>");
print("Options:");
print(" -f STR output format: aln (BLAST-like), maf or lastz-cigar [aln]");
print(" -l INT line length in BLAST-like output [80]");
exit(1);
}
function padding_str(x, len, right)
{
var s = x.toString();
if (s.length < len) {
if (right) s += Array(len - s.length + 1).join(" ");
else s = Array(len - s.length + 1).join(" ") + s;
}
return s;
}
function update_aln(s_ref, s_qry, s_mid, type, seq, slen)
{
var l = type == '*'? 1 : seq.length;
if (type == '=' || type == ':') {
s_ref.set(seq);
s_qry.set(seq);
s_mid.set(Array(l+1).join("|"));
slen[0] += l, slen[1] += l;
} else if (type == '*') {
s_ref.set(seq.charAt(0));
s_qry.set(seq.charAt(1));
s_mid.set(' ');
slen[0] += 1, slen[1] += 1;
} else if (type == '+') {
s_ref.set(Array(l+1).join("-"));
s_qry.set(seq);
s_mid.set(Array(l+1).join(" "));
slen[1] += l;
} else if (type == '-') {
s_ref.set(seq);
s_qry.set(Array(l+1).join("-"));
s_mid.set(Array(l+1).join(" "));
slen[0] += l;
}
}
function print_aln(rs, qs, strand, slen, elen, s_ref, s_qry, s_mid)
{
print(["Ref+:", padding_str(rs + slen[0] + 1, 10, false), s_ref.toString(), padding_str(rs + elen[0], 10, true)].join(" "));
print(" " + s_mid.toString());
var st, en;
if (strand == '+') st = qs + slen[1] + 1, en = qs + elen[1];
else st = qs - slen[1], en = qs - elen[1] + 1;
print(["Qry" + strand + ":", padding_str(st, 10, false), s_qry.toString(), padding_str(en, 10, true)].join(" "));
}
var s_ref = new Bytes(), s_qry = new Bytes(), s_mid = new Bytes(); // these are used to show padded alignment
var re_cs = /([:=\-\+\*])(\d+|[A-Za-z]+)/g;
var re_cg = /(\d+)([MIDNSH])/g;
var buf = new Bytes();
var file = args[getopt.ind] == "-"? new File() : new File(args[getopt.ind]);
var lineno = 0;
if (fmt == "maf") print("##maf version=1\n");
while (file.readline(buf) >= 0) {
var m, line = buf.toString();
var t = line.split("\t", 12);
++lineno;
s_ref.length = s_qry.length = s_mid.length = 0;
var slen = [0, 0], elen = [0, 0];
if (fmt == "lastz-cigar") { // LASTZ-cigar output
var cg = (m = /\tcg:Z:(\S+)/.exec(line)) != null? m[1] : null;
if (cg == null) {
warn("WARNING: converting to LASTZ-cigar format requires the 'cg' tag, which is absent on line " + lineno);
continue;
}
var score = (m = /\tAS:i:(\d+)/.exec(line)) != null? m[1] : 0;
var out = ['cigar:', t[0], t[2], t[3], t[4], t[5], t[7], t[8], '+', score];
while ((m = re_cg.exec(cg)) != null)
out.push(m[2], m[1]);
print(out.join(" "));
} else if (fmt == "maf") { // MAF output
var cs = (m = /\tcs:Z:(\S+)/.exec(line)) != null? m[1] : null;
if (cs == null) {
warn("WARNING: converting to MAF requires the 'cs' tag, which is absent on line " + lineno);
continue;
}
while ((m = re_cs.exec(cs)) != null) {
if (m[1] == ':')
throw Error("converting to MAF only works with 'minimap2 --cs=long'");
update_aln(s_ref, s_qry, s_mid, m[1], m[2], elen);
}
var score = (m = /\tAS:i:(\d+)/.exec(line)) != null? parseInt(m[1]) : 0;
var len = t[0].length > t[5].length? t[0].length : t[5].length;
print("a " + score);
print(["s", padding_str(t[5], len, true), padding_str(t[7], 10, false), padding_str(parseInt(t[8]) - parseInt(t[7]), 10, false),
"+", padding_str(t[6], 10, false), s_ref.toString()].join(" "));
var qs, qe, ql = parseInt(t[1]);
if (t[4] == '+') {
qs = parseInt(t[2]);
qe = parseInt(t[3]);
} else {
qs = ql - parseInt(t[3]);
qe = ql - parseInt(t[2]);
}
print(["s", padding_str(t[0], len, true), padding_str(qs, 10, false), padding_str(qe - qs, 10, false),
t[4], padding_str(ql, 10, false), s_qry.toString()].join(" "));
print("");
} else { // BLAST-like output
var cs = (m = /\tcs:Z:(\S+)/.exec(line)) != null? m[1] : null;
if (cs == null) {
warn("WARNING: converting to BLAST-like alignment requires the 'cs' tag, which is absent on line " + lineno);
continue;
}
line = line.replace(/\tc[sg]:Z:\S+/g, ""); // get rid of cs or cg tags
print('>' + line);
var rs = parseInt(t[7]), qs = t[4] == '+'? parseInt(t[2]) : parseInt(t[3]);
var n_blocks = 0;
while ((m = re_cs.exec(cs)) != null) {
if (m[1] == ':') m[2] = Array(parseInt(m[2]) + 1).join("=");
var start = 0, rest = m[1] == '*'? 1 : m[2].length;
while (rest > 0) {
var l_proc;
if (s_ref.length + rest >= line_len) {
l_proc = line_len - s_ref.length;
update_aln(s_ref, s_qry, s_mid, m[1], m[1] == '*'? m[2] : m[2].substr(start, l_proc), elen);
if (n_blocks > 0) print("");
print_aln(rs, qs, t[4], slen, elen, s_ref, s_qry, s_mid);
++n_blocks;
s_ref.length = s_qry.length = s_mid.length = 0;
slen[0] = elen[0], slen[1] = elen[1];
} else {
l_proc = rest;
update_aln(s_ref, s_qry, s_mid, m[1], m[1] == '*'? m[2] : m[2].substr(start, l_proc), elen);
}
rest -= l_proc, start += l_proc;
}
}
if (s_ref.length > 0) {
if (n_blocks > 0) print("");
print_aln(rs, qs, t[4], slen, elen, s_ref, s_qry, s_mid);
++n_blocks;
}
print("//");
}
}
file.close();
buf.destroy();
s_ref.destroy(); s_qry.destroy(); s_mid.destroy();
}
function paf_gff2bed(args)
{
var c, fn_ucsc_fai = null, is_short = false, keep_gff = false, print_junc = false;
while ((c = getopt(args, "u:sgj")) != null) {
if (c == 'u') fn_ucsc_fai = getopt.arg;
else if (c == 's') is_short = true;
else if (c == 'g') keep_gff = true;
else if (c == 'j') print_junc = true;
}
if (getopt.ind == args.length) {
print("Usage: paftools.js gff2bed [options] <in.gff>");
print("Options:");
print(" -j Output junction BED");
print(" -s Print names in the short form");
print(" -u FILE hg38.fa.fai for chr name conversion");
print(" -g Output GFF (used with -u)");
exit(1);
}
var ens2ucsc = {};
if (fn_ucsc_fai != null) {
var buf = new Bytes();
var file = new File(fn_ucsc_fai);
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
var s = t[0];
if (/_(random|alt|decoy)$/.test(s)) {
s = s.replace(/_(random|alt|decoy)$/, '');
s = s.replace(/^chr\S+_/, '');
} else {
s = s.replace(/^chrUn_/, '');
}
s = s.replace(/v(\d+)/, ".$1");
if (s != t[0]) ens2ucsc[s] = t[0];
}
file.close();
buf.destroy();
}
var colors = {
'protein_coding':'0,128,255',
'lincRNA':'0,192,0',
'snRNA':'0,192,0',
'miRNA':'0,192,0',
'misc_RNA':'0,192,0'
};
function print_bed12(exons, cds_st, cds_en, is_short, print_junc)
{
if (exons.length == 0) return;
var name = is_short? exons[0][7] + "|" + exons[0][5] : exons[0].slice(4, 7).join("|");
var a = exons.sort(function(a,b) {return a[1]-b[1]});
if (print_junc) {
for (var i = 1; i < a.length; ++i)
print(a[i][0], a[i-1][2], a[i][1], name, 1000, a[i][3]);
return;
}
var sizes = [], starts = [], st, en;
st = a[0][1];
en = a[a.length - 1][2];
if (cds_st == 1<<30) cds_st = st;
if (cds_en == 0) cds_en = en;
if (cds_st < st || cds_en > en)
throw Error("inconsistent thick start or end for transcript " + a[0][4]);
for (var i = 0; i < a.length; ++i) {
sizes.push(a[i][2] - a[i][1]);
starts.push(a[i][1] - st);
}
var color = colors[a[0][5]];
if (color == null) color = '196,196,196';
print(a[0][0], st, en, name, 1000, a[0][3], cds_st, cds_en, color, a.length, sizes.join(",") + ",", starts.join(",") + ",");
}
var re_gtf = /(transcript_id|transcript_type|transcript_biotype|gene_name|transcript_name) "([^"]+)";/g;
var re_gff3 = /(transcript_id|transcript_type|transcript_biotype|gene_name|transcript_name)=([^;]+)/g;
var buf = new Bytes();
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var exons = [], cds_st = 1<<30, cds_en = 0, last_id = null;
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
if (keep_gff) {
if (t[0].charAt(0) != '#' && ens2ucsc[t[0]] != null)
t[0] = ens2ucsc[t[0]];
print(t.join("\t"));
continue;
}
if (t[0].charAt(0) == '#') continue;
if (t[2] != "CDS" && t[2] != "exon") continue;
t[3] = parseInt(t[3]) - 1;
t[4] = parseInt(t[4]);
var id = null, type = "", gname = "N/A", biotype = "", m, tname = "N/A";
while ((m = re_gtf.exec(t[8])) != null) {
if (m[1] == "transcript_id") id = m[2];
else if (m[1] == "transcript_type") type = m[2];
else if (m[1] == "transcript_biotype") biotype = m[2];
else if (m[1] == "gene_name") name = m[2];
else if (m[1] == "transcript_name") tname = m[2];
}
while ((m = re_gff3.exec(t[8])) != null) {
if (m[1] == "transcript_id") id = m[2];
else if (m[1] == "transcript_type") type = m[2];
else if (m[1] == "transcript_biotype") biotype = m[2];
else if (m[1] == "gene_name") name = m[2];
else if (m[1] == "transcript_name") tname = m[2];
}
if (type == "" && biotype != "") type = biotype;
if (id == null) throw Error("No transcript_id");
if (id != last_id) {
print_bed12(exons, cds_st, cds_en, is_short, print_junc);
exons = [], cds_st = 1<<30, cds_en = 0;
last_id = id;
}
if (t[2] == "CDS") {
cds_st = cds_st < t[3]? cds_st : t[3];
cds_en = cds_en > t[4]? cds_en : t[4];
} else if (t[2] == "exon") {
if (fn_ucsc_fai != null) {
if (ens2ucsc[t[0]] != null)
t[0] = ens2ucsc[t[0]];
else if (/^[A-Z]+\d+\.\d+$/.test(t[0]))
t[0] = t[0].replace(/([A-Z]+\d+)\.(\d+)/, "chrUn_$1v$2");
}
exons.push([t[0], t[3], t[4], t[6], id, type, name, tname]);
}
}
if (last_id != null)
print_bed12(exons, cds_st, cds_en, is_short, print_junc);
file.close();
buf.destroy();
}
function paf_sam2paf(args)
{
var c, pri_only = false, long_cs = false;
while ((c = getopt(args, "pL")) != null) {
if (c == 'p') pri_only = true;
else if (c == 'L') long_cs = true;
}
if (args.length == getopt.ind) {
print("Usage: paftools.js sam2paf [options] <in.sam>");
print("Options:");
print(" -p convert primary or supplementary alignments only");
print(" -L output the cs tag in the long form");
exit(1);
}
var file = args[getopt.ind] == "-"? new File() : new File(args[getopt.ind]);
var buf = new Bytes();
var re = /(\d+)([MIDSHNX=])/g, re_MD = /(\d+)|(\^[A-Za-z]+)|([A-Za-z])/g, re_tag = /\t(\S\S:[AZif]):(\S+)/g;
var ctg_len = {}, lineno = 0;
while (file.readline(buf) >= 0) {
var m, n_cigar = 0, line = buf.toString();
++lineno;
if (line.charAt(0) == '@') {
if (/^@SQ/.test(line)) {
var name = (m = /\tSN:(\S+)/.exec(line)) != null? m[1] : null;
var l = (m = /\tLN:(\d+)/.exec(line)) != null? parseInt(m[1]) : null;
if (name != null && l != null) ctg_len[name] = l;
}
continue;
}
var t = line.split("\t", 11);
var flag = parseInt(t[1]);
if (t[9] != '*' && t[10] != '*' && t[9].length != t[10].length)
throw Error("at line " + lineno + ": inconsistent SEQ and QUAL lengths - " + t[9].length + " != " + t[10].length);
if (t[2] == '*' || (flag&4) || t[5] == '*') continue;
if (pri_only && (flag&0x100)) continue;
var tlen = ctg_len[t[2]];
if (tlen == null) throw Error("at line " + lineno + ": can't find the length of contig " + t[2]);
// find tags
var nn = 0, NM = null, MD = null, cs_str = null, md_list = [];
while ((m = re_tag.exec(line)) != null) {
if (m[1] == "NM:i") NM = parseInt(m[2]);
else if (m[1] == "nn:i") nn = parseInt(m[2]);
else if (m[1] == "MD:Z") MD = m[2];
else if (m[1] == "cs:Z") cs_str = m[2];
}
if (t[9] == '*') MD = cs_str = null;
// infer various lengths from CIGAR
var clip = [0, 0], soft_clip = 0, I = [0, 0], D = [0, 0], M = 0, N = 0, mm = 0, have_M = false, have_ext = false, cigar = [];
while ((m = re.exec(t[5])) != null) {
var l = parseInt(m[1]), op = m[2];
if (op == 'M') M += l, have_M = true;
else if (op == 'I') ++I[0], I[1] += l;
else if (op == 'D') ++D[0], D[1] += l;
else if (op == 'N') N += l;
else if (op == 'S') clip[n_cigar == 0? 0 : 1] = l, soft_clip += l;
else if (op == 'H') clip[n_cigar == 0? 0 : 1] = l;
else if (op == '=') M += l, have_ext = true, op = 'M';
else if (op == 'X') M += l, mm += l, have_ext = true, op = 'M';
++n_cigar;
if (MD != null && op != 'H') {
if (cigar.length > 0 && cigar[cigar.length-1][1] == op)
cigar[cigar.length-1][0] += l;
else cigar.push([l, op]);
}
}
var ql = M + I[1] + soft_clip;
var tl = M + D[1] + N;
var ts = parseInt(t[3]) - 1, te = ts + tl;
// checking coordinate and length consistencies
if (n_cigar > 65535)
warn("WARNING at line " + lineno + ": " + n_cigar + " CIGAR operations");
if (te > tlen) {
warn("WARNING at line " + lineno + ": alignment end position larger than ref length; skipped");
continue;
}
if (t[9] != '*' && t[9].length != ql) {
warn("WARNING at line " + lineno + ": SEQ length inconsistent with CIGAR (" + t[9].length + " != " + ql + "); skipped");
continue;
}
// parse MD
var cs = [];
if (MD != null && cs_str == null && t[9] != "*") {
var k = 0, cx = 0, cy = 0, mx = 0, my = 0; // cx: cigar ref position; cy: cigar query; mx: MD ref; my: MD query
while ((m = re_MD.exec(MD)) != null) {
if (m[2] != null) { // deletion from the reference
var len = m[2].length - 1;
cs.push('-', m[2].substr(1));
mx += len, cx += len, ++k;
} else { // copy or mismatch
var ml = m[1] != null? parseInt(m[1]) : 1;
while (k < cigar.length && cigar[k][1] != 'D') {
var cl = cigar[k][0], op = cigar[k][1];
if (op == 'M') {
if (my + ml < cy + cl) {
if (ml > 0) {
if (m[3] != null) cs.push('*', m[3], t[9][my]);
else if (long_cs) cs.push('=', t[9].substr(my, ml));
else cs.push(':', ml);
}
mx += ml, my += ml, ml = 0;
break;
} else {
var dl = cy + cl - my;
if (long_cs) cs.push('=', t[9].substr(my, dl));
else cs.push(':', dl);
cx += cl, cy += cl, ++k;
mx += dl, my += dl, ml -= dl;
}
} else if (op == 'I') {
cs.push('+', t[9].substr(cy, cl));
cy += cl, my += cl, ++k;
} else if (op == 'S') {
cy += cl, my += cl, ++k;
} else throw Error("at line " + lineno + ": inconsistent MD tag");
}
if (ml != 0) throw Error("at line " + lineno + ": inconsistent MD tag");
}
}
if (cx != mx || cy != my) throw Error("at line " + lineno + ": inconsistent MD tag");
}
// compute matching length, block length and calibrate NM
if (have_ext && !have_M) { // extended CIGAR
if (NM != null && NM != I[1] + D[1] + mm)
warn("WARNING at line " + lineno + ": NM is different from sum of gaps and mismatches");
NM = I[1] + D[1] + mm;
} else if (NM != null) { // standard CIGAR; NM present
if (NM < I[1] + D[1]) {
warn("WARNING at line " + lineno + ": NM is less than the total number of gaps (" + NM + " < " + (I[1]+D[1]) + ")");
NM = I[1] + D[1];
}
mm = NM - (I[1] + D[1]);
} else { // no way to compute mm
warn("WARNING at line " + lineno + ": unable to find the number of mismatches; assuming zero");
mm = 0;
}
var mlen = M - mm;
var blen = M + I[1] + D[1];
// find query name, start and end
var qlen = M + I[1] + clip[0] + clip[1];
var qname = t[0], qs, qe;
if ((flag&1) && (flag&0x40)) qname += '/1';
if ((flag&1) && (flag&0x80)) qname += '/2';
if (flag&16) qs = clip[1], qe = qlen - clip[0];
else qs = clip[0], qe = qlen - clip[1];
// optional tags
var type = flag&0x100? 'S' : 'P';
var tags = ["tp:A:" + type];
if (NM != null) tags.push("mm:i:"+mm);
tags.push("gn:i:"+(I[1]+D[1]), "go:i:"+(I[0]+D[0]), "cg:Z:" + t[5].replace(/\d+[SH]/g, ''));
if (cs_str != null) tags.push("cs:Z:" + cs_str);
else if (cs.length > 0) tags.push("cs:Z:" + cs.join(""));
// print out
var a = [qname, qlen, qs, qe, flag&16? '-' : '+', t[2], tlen, ts, te, mlen, blen, t[4]];
print(a.join("\t"), tags.join("\t"));
}
buf.destroy();
file.close();
}
function paf_delta2paf(args)
{
if (args.length == 0) {
print("Usage: paftools.js delta2paf <in.delta>");
exit(1);
}
var buf = new Bytes();
var file = args[0] == '-'? new File() : new File(args[0]);
var rname, qname, rlen, qlen, qs, qe, rs, re, strand, NM, cigar, x, y, seen_gt = false;
while (file.readline(buf) >= 0) {
var m, line = buf.toString();
if ((m = /^>(\S+)\s+(\S+)\s+(\d+)\s+(\d+)/.exec(line)) != null) {
rname = m[1], qname = m[2], rlen = parseInt(m[3]), qlen = parseInt(m[4]);
seen_gt = true;
continue;
}
if (!seen_gt) continue;
var t = line.split(" ");
if (t.length == 7) {
for (var i = 0; i < 5; ++i)
t[i] = parseInt(t[i]);
strand = ((t[0] < t[1] && t[2] < t[3]) || (t[0] > t[1] && t[2] > t[3]))? 1 : -1;
rs = (t[0] < t[1]? t[0] : t[1]) - 1;
re = t[1] > t[0]? t[1] : t[0];
qs = (t[2] < t[3]? t[2] : t[3]) - 1;
qe = t[3] > t[2]? t[3] : t[2];
x = y = 0;
NM = parseInt(t[4]);
cigar = [];
} else if (t.length == 1) {
var d = parseInt(t[0]);
if (d == 0) {
var blen = 0, cigar_str = [];
if (re - rs - x != qe - qs - y) throw Error("inconsisnt alignment");
cigar.push((re - rs - x) << 4);
for (var i = 0; i < cigar.length; ++i) {
blen += cigar[i] >> 4;
cigar_str.push((cigar[i]>>4) + "MID".charAt(cigar[i]&0xf));
}
print([qname, qlen, qs, qe, strand > 0? '+' : '-', rname, rlen, rs, re, blen - NM, blen, 0, "NM:i:" + NM, "cg:Z:" + cigar_str.join("")].join("\t"));
} else if (d > 0) {
var l = d - 1;
x += l + 1, y += l;
if (l > 0) cigar.push(l<<4);
if (cigar.length > 0 && (cigar[cigar.length-1]&0xf) == 2)
cigar[cigar.length-1] += 1<<4;
else cigar.push(1<<4|2); // deletion
} else {
var l = -d - 1;
x += l, y += l + 1;
if (l > 0) cigar.push(l<<4);
if (cigar.length > 0 && (cigar[cigar.length-1]&0xf) == 1)
cigar[cigar.length-1] += 1<<4;
else cigar.push(1<<4|1); // insertion
}
}
}
file.close();
buf.destroy();
}
function paf_splice2bed(args)
{
var colors = ["0,128,255", "255,0,0", "0,192,0"];
function print_lines(a, fmt, keep_multi)
{
if (a.length == 0) return;
if (fmt == "bed") {
var n_pri = 0;
for (var i = 0; i < a.length; ++i)
if (a[i][8] == 0) ++n_pri;
if (n_pri > 1) {
for (var i = 0; i < a.length; ++i)
if (a[i][8] == 0) a[i][8] = 1;
} else if (n_pri == 0) {
warn("Warning: " + a[0][3] + " doesn't have a primary alignment");
}
for (var i = 0; i < a.length; ++i) {
if (!keep_multi && a[i][8] == 2) continue;
a[i][8] = colors[a[i][8]];
print(a[i].join("\t"));
}
}
a.length = 0;
}
var re = /(\d+)([MIDNSH])/g;
var c, fmt = "bed", fn_name_conv = null, keep_multi = false;
while ((c = getopt(args, "f:n:m")) != null) {
if (c == 'f') fmt = getopt.arg;
else if (c == 'n') fn_name_conv = getopt.arg;
else if (c == 'm') keep_multi = true;
}
if (getopt.ind == args.length) {
print("Usage: paftools.js splice2bed [options] <in.paf>|<in.sam>");
print("Options:");
print(" -m keep multiple mappings (SAM flag 0x100)");
exit(1);
}
var conv = null;
if (fn_name_conv != null) {
conv = new Map();
var file = new File(fn_name_conv);
var buf = new Bytes();
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
conv.put(t[0], t[1]);
}
buf.destroy();
file.close();
}
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var buf = new Bytes();
var a = [];
while (file.readline(buf) >= 0) {
var line = buf.toString();
if (line.charAt(0) == '@') continue; // skip SAM header lines
var t = line.split("\t");
var is_pri = false, cigar = null, a1;
var qname = conv != null? conv.get(t[0]) : null;
if (qname != null) t[0] = qname;
if (t.length >= 10 && t[4] != '+' && t[4] != '-' && /^\d+/.test(t[1])) { // SAM
var flag = parseInt(t[1]);
if (flag&1) t[0] += '/' + (flag>>6&3);
}
if (a.length && a[0][3] != t[0]) {
print_lines(a, fmt, keep_multi);
a = [];
}
if (t.length >= 12 && (t[4] == '+' || t[4] == '-')) { // PAF
for (var i = 12; i < t.length; ++i) {
if (t[i].substr(0, 5) == 'cg:Z:') {
cigar = t[i].substr(5);
} else if (t[i].substr(0, 5) == 's2:i:') {
is_pri = true;
}
}
a1 = [t[5], t[7], t[8], t[0], Math.floor(t[9]/t[10]*1000), t[4]];
} else if (t.length >= 10) { // SAM
var flag = parseInt(t[1]);
if ((flag&4) || a[2] == '*') continue;
cigar = t[5];
is_pri = (flag&0x100)? false : true;
a1 = [t[2], parseInt(t[3])-1, null, t[0], 1000, (flag&16)? '-' : '+'];
} else {
throw Error("unrecognized input format");
}
if (cigar == null) throw Error("missing CIGAR");
var m, x0 = 0, x = 0, bs = [], bl = [];
while ((m = re.exec(cigar)) != null) {
if (m[2] == 'M' || m[2] == 'D') {
x += parseInt(m[1]);
} else if (m[2] == 'N') {
bs.push(x0);
bl.push(x - x0);
x += parseInt(m[1]);
x0 = x;
}
}
bs.push(x0);
bl.push(x - x0);
// write the BED12 line
if (a1[2] == null) a1[2] = a1[1] + x;
a1.push(a1[1], a1[2]); // thick start/end is the same as start/end
a1.push(is_pri? 0 : 2, bs.length, bl.join(",")+",", bs.join(",")+",");
a.push(a1);
}
print_lines(a, fmt, keep_multi);
buf.destroy();
file.close();
if (conv != null) conv.destroy();
}
/**********************
* Evaluation related *
**********************/
// evaluate mapping accuracy
function paf_mapeval(args)
{
var c, max_mapq = 60, mode = 0, err_out_q = 256, print_err = false, ovlp_ratio = 0.1, cap_short_mapq = false;
while ((c = getopt(args, "Q:r:m:c")) != null) {
if (c == 'Q') err_out_q = parseInt(getopt.arg), print_err = true;
else if (c == 'r') ovlp_ratio = parseFloat(getopt.arg);
else if (c == 'm') mode = parseInt(getopt.arg);
else if (c == 'c') cap_short_mapq = true;
}
if (args.length == getopt.ind) {
warn("Usage: paftools.js mapeval [options] <in.paf>|<in.sam>");
warn("Options:");
warn(" -r FLOAT mapping correct if overlap_length/union_length>FLOAT [" + ovlp_ratio + "]");
warn(" -Q INT print wrong mappings with mapQ>INT [don't print]");
warn(" -m INT 0: eval the longest aln only; 1: first aln only; 2: all primary aln [0]");
exit(1);
}
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var buf = new Bytes();
var tot = [], err = [];
for (var q = 0; q <= max_mapq; ++q)
tot[q] = err[q] = 0;
function is_correct(s, b)
{
if (s[0] != b[0] || s[3] != b[3]) return false;
var o, l;
if (s[1] < b[1]) {
if (s[2] <= b[1]) return false;
o = (s[2] < b[2]? s[2] : b[2]) - b[1];
l = (s[2] > b[2]? s[2] : b[2]) - s[1];
} else {
if (b[2] <= s[1]) return false;
o = (s[2] < b[2]? s[2] : b[2]) - s[1];
l = (s[2] > b[2]? s[2] : b[2]) - b[1];
}
return o/l > ovlp_ratio? true : false;
}
function count_err(qname, a, tot, err, mode)
{
if (a.length == 0) return;
var m, s;
if ((m = /^(\S+)!(\S+)!(\d+)!(\d+)!([\+\-])$/.exec(qname)) != null) { // pbsim single-end reads
s = [m[1], m[2], parseInt(m[3]), parseInt(m[4]), m[5]];
} else if ((m = /^(\S+)!(\S+)!(\d+)_(\d+)!(\d+)_(\d+)!([\+\-])([\+\-])\/([12])$/.exec(qname)) != null) { // mason2 paired-end reads
if (m[9] == '1') {
s = [m[1], m[2], parseInt(m[3]), parseInt(m[5]), m[7]];
} else {
s = [m[1], m[2], parseInt(m[4]), parseInt(m[6]), m[8]];
}
} else throw Error("Failed to parse simulated read names '" + qname + "'");
s.shift(); // skip the orginal read name
if (mode == 0 || mode == 1) { // longest only or first only
var max_i = 0;
if (mode == 0) { // longest only
var max = 0;
for (var i = 0; i < a.length; ++i)
if (a[i][5] > max)
max = a[i][5], max_i = i;
}
var mapq = a[max_i][4];
++tot[mapq];
if (!is_correct(s, a[max_i])) {
if (mapq >= err_out_q)
print('E', qname, a[max_i].join("\t"));
++err[mapq];
}
} else if (mode == 2) { // all primary mode
var max_err_mapq = -1, max_mapq = 0, max_err_i = -1;
if (cap_short_mapq) {
var max = 0, max_q = 0;
for (var i = 0; i < a.length; ++i)
if (a[i][5] > max)
max = a[i][5], max_q = a[i][4];
for (var i = 0; i < a.length; ++i)
a[i][4] = max_q < a[i][4]? max_q : a[i][4];
}
for (var i = 0; i < a.length; ++i) {
max_mapq = max_mapq > a[i][4]? max_mapq : a[i][4];
if (!is_correct(s, a[i]))
if (a[i][4] > max_err_mapq)
max_err_mapq = a[i][4], max_err_i = i;
}
if (max_err_mapq >= 0) {
++tot[max_err_mapq], ++err[max_err_mapq];
if (max_err_mapq >= err_out_q)
print('E', qname, a[max_err_i].join("\t"));
} else ++tot[max_mapq];
}
}
var lineno = 0, last = null, a = [], n_unmapped = null;
var re_cigar = /(\d+)([MIDSHN])/g;
while (file.readline(buf) >= 0) {
var m, line = buf.toString();
++lineno;
if (line[0] != '@') {
var t = line.split("\t");
if (t[4] == '+' || t[4] == '-') { // PAF
if (last != t[0]) {
if (last != null) count_err(last, a, tot, err, mode);
a = [], last = t[0];
}
if (/\ts1:i:\d+/.test(line) && !/\ts2:i:\d+/.test(line)) // secondary alignment in minimap2 PAF
continue;
var mapq = parseInt(t[11]);
if (mapq > max_mapq) mapq = max_mapq;
a.push([t[5], parseInt(t[7]), parseInt(t[8]), t[4], mapq, parseInt(t[9])]);
} else { // SAM
var flag = parseInt(t[1]);
var read_no = flag>>6&0x3;
var qname = t[0];
if (!/\/[12]$/.test(qname))
qname = read_no == 1 || read_no == 2? t[0] + '/' + read_no : t[0];
if (last != qname) {
if (last != null) count_err(last, a, tot, err, mode);
a = [], last = qname;
}
if (flag&0x100) continue; // secondary alignment
if ((flag&0x4) || t[2] == '*') { // unmapped
if (n_unmapped == null) n_unmapped = 0;
++n_unmapped;
continue;
}
var mapq = parseInt(t[4]);
if (mapq > max_mapq) mapq = max_mapq;
var pos = parseInt(t[3]) - 1, pos_end = pos;
var n_gap = 0, mlen = 0;
while ((m = re_cigar.exec(t[5])) != null) {
var len = parseInt(m[1]);
if (m[2] == 'M') pos_end += len, mlen += len;
else if (m[2] == 'I') n_gap += len;
else if (m[2] == 'D') n_gap += len, pos_end += len;
}
var score = pos_end - pos;
if ((m = /\tNM:i:(\d+)/.exec(line)) != null) {
var NM = parseInt(m[1]);
if (NM >= n_gap) score = mlen - (NM - n_gap);
}
a.push([t[2], pos, pos_end, (flag&16)? '-' : '+', mapq, score]);
}
}
}
if (last != null) count_err(last, a, tot, err, mode);
buf.destroy();
file.close();
var sum_tot = 0, sum_err = 0, q_out = -1, sum_tot2 = 0, sum_err2 = 0;
for (var q = max_mapq; q >= 0; --q) {
if (tot[q] == 0) continue;
if (q_out < 0 || err[q] > 0) {
if (q_out >= 0) print('Q', q_out, sum_tot, sum_err, (sum_err2/sum_tot2).toFixed(9), sum_tot2);
sum_tot = sum_err = 0, q_out = q;
}
sum_tot += tot[q], sum_err += err[q];
sum_tot2 += tot[q], sum_err2 += err[q];
}
print('Q', q_out, sum_tot, sum_err, (sum_err2/sum_tot2).toFixed(9), sum_tot2);
if (n_unmapped != null) print('U', n_unmapped);
}
// convert mason2 SAM to FASTQ
function paf_mason2fq(args)
{
if (args.length == 0) {
print("Usage: paftools.js mason2fq <mason.sam>");
exit(1);
}
function print_se(a)
{
print('@' + a.slice(0, 5).join("!") + " " + a[8]);
print(a[5]);
print("+");
print(a[6]);
}
var buf = new Bytes(), buf2 = new Bytes();
var file = new File(args[0]);
var re = /(\d+)([MIDSHN])/g;
var last = null;
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
if (t[0].charAt(0) == '@') continue;
var m, l_ref = 0;
while ((m = re.exec(t[5])) != null)
if (m[2] == 'D' || m[2] == 'M' || m[2] == 'N')
l_ref += parseInt(m[1]);
var flag = parseInt(t[1]);
var rev = !!(flag&16);
var seq, qual;
if (rev) {
buf2.length = 0;
buf2.set(t[9], 0);
buf2.revcomp();
seq = buf2.toString();
buf2.set(t[10], 0);
buf2.reverse();
qual = buf2.toString();
} else seq = t[9], qual = t[10];
var qname = t[0];
qname = qname.replace(/^simulated./, "");
var chr = t[2];
var pos = parseInt(t[3]) - 1;
var strand = (flag&16)? '-' : '+';
var read_no = flag&0xc0;
if (read_no == 0x40) read_no = 1;
else if (read_no == 0x80) read_no = 2;
else read_no = 0;
var err = 0, snp = 0, indel = 0;
for (var i = 11; i < t.length; ++i) {
if ((m = /^XE:i:(\d+)/.exec(t[i])) != null) err = m[1];
else if ((m = /^XS:i:(\d+)/.exec(t[i])) != null) snp = m[1];
else if ((m = /^XI:i:(\d+)/.exec(t[i])) != null) indel = m[1];
}
var comment = [err, snp, indel].join(":");
if (last == null) {
last = [qname, chr, pos, pos + l_ref, strand, seq, qual, read_no, comment];
} else if (last[0] != qname) {
print_se(last);
last = [qname, chr, pos, pos + l_ref, strand, seq, qual, read_no, comment];
} else {
if (read_no == 2) { // last[] is the first read
if (last[7] != 1) throw Error("ERROR: can't find read1");
var name = [qname, chr, last[2] + "_" + pos, last[3] + "_" + (pos + l_ref), last[4] + strand].join("!");
print('@' + name + '/1' + ' ' + last[8]); print(last[5]); print("+"); print(last[6]);
print('@' + name + '/2' + ' ' + comment); print(seq); print("+"); print(qual);
} else {
if (last[7] != 2) throw Error("ERROR: can't find read2");
var name = [qname, chr, pos + "_" + last[2], (pos + l_ref) + "_" + last[3], strand + last[4]].join("!");
print('@' + name + '/1' + ' ' + comment); print(seq); print("+"); print(qual);
print('@' + name + '/2' + ' ' + last[8]); print(last[5]); print("+"); print(last[6]);
}
last = null;
}
}
if (last != null) print_se(last);
file.close();
buf.destroy();
buf2.destroy();
}
// convert pbsim MAF to FASTQ
function paf_pbsim2fq(args)
{
if (args.length < 2) {
print("Usage: paftools.js pbsim2fq <ref.fa.fai> <pbsim1.maf> [[pbsim2.maf] ...]");
exit(1);
}
var file, buf = new Bytes(), buf2 = new Bytes();
file = new File(args[0]);
var chr_list = [];
while (file.readline(buf) >= 0) {
var t = buf.toString().split(/\s+/);
chr_list.push(t[0]);
}
file.close();
for (var k = 1; k < args.length; ++k) {
var fn = args[k];
file = new File(fn);
var state = 0, reg;
while (file.readline(buf) >= 0) {
var line = buf.toString();
if (state == 0 && line.charAt(0) == 'a') {
state = 1;
} else if (state == 1 && line.charAt(0) == 's') {
var t = line.split(/\s+/);
var st = parseInt(t[2]);
reg = [st, st + parseInt(t[3])];
state = 2;
} else if (state == 2 && line.charAt(0) == 's') {
var m, t = line.split(/\s+/);
if ((m = /S(\d+)_\d+/.exec(t[1])) == null) throw Error("Failed to parse the read name");
var chr_id = parseInt(m[1]) - 1;
if (chr_id >= chr_list.length) throw Error("Index outside the chr list");
var name = [t[1], chr_list[chr_id], reg[0], reg[1], t[4]].join("!");
var seq = t[6].replace(/\-/g, "");
if (seq.length != parseInt(t[5])) throw Error("Inconsistent read length");
if (seq.indexOf("NN") < 0) {
if (t[4] == '-') {
buf2.set(seq, 0);
buf2.length = seq.length;
buf2.revcomp();
seq = buf2.toString();
}
print(">" + name);
print(seq);
}
state = 0;
}
}
file.close();
}
buf.destroy();
buf2.destroy();
}
function paf_junceval(args)
{
var c, l_fuzzy = 0, print_ovlp = false, print_err_only = false, first_only = false, chr_only = false;
while ((c = getopt(args, "l:epc")) != null) {
if (c == 'l') l_fuzzy = parseInt(getopt.arg);
else if (c == 'e') print_err_only = print_ovlp = true;
else if (c == 'p') print_ovlp = true;
else if (c == 'c') chr_only = true;
}
if (args.length - getopt.ind < 1) {
print("Usage: paftools.js junceval [options] <gene.gtf> <aln.sam>");
print("Options:");
print(" -l INT tolerance of junction positions (0 for exact) [0]");
print(" -p print overlapping introns");
print(" -e print erroreous overlapping introns");
print(" -c only consider alignments to /^(chr)?([0-9]+|X|Y)$/");
exit(1);
}
var file, buf = new Bytes();
var tr = {};
file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
while (file.readline(buf) >= 0) {
var m, t = buf.toString().split("\t");
if (t[0].charAt(0) == '#') continue;
if (t[2] != 'exon') continue;
var st = parseInt(t[3]) - 1;
var en = parseInt(t[4]);
if ((m = /transcript_id "(\S+)"/.exec(t[8])) == null) continue;
var tid = m[1];
if (tr[tid] == null) tr[tid] = [t[0], t[6], 0, 0, []];
tr[tid][4].push([st, en]);
}
file.close();
var anno = {};
for (var tid in tr) {
var t = tr[tid];
Interval.sort(t[4]);
t[2] = t[4][0][0];
t[3] = t[4][t[4].length - 1][1];
if (anno[t[0]] == null) anno[t[0]] = [];
var s = t[4];
for (var i = 0; i < s.length - 1; ++i) {
if (s[i][1] >= s[i+1][0])
warn("WARNING: incorrect annotation for transcript "+tid+" ("+s[i][1]+" >= "+s[i+1][0]+")")
anno[t[0]].push([s[i][1], s[i+1][0]]);
}
}
tr = null;
for (var chr in anno) {
var e = anno[chr];
if (e.length == 0) continue;
Interval.sort(e);
var k = 0;
for (var i = 1; i < e.length; ++i) // dedup
if (e[i][0] != e[k][0] || e[i][1] != e[k][1])
e[++k] = e[i].slice(0);
e.length = k + 1;
Interval.index_end(e);
}
var n_pri = 0, n_unmapped = 0, n_mapped = 0;
var n_sgl = 0, n_splice = 0, n_splice_hit = 0, n_splice_novel = 0;
file = getopt.ind+1 >= args.length || args[getopt.ind+1] == '-'? new File() : new File(args[getopt.ind+1]);
var last_qname = null;
var re_cigar = /(\d+)([MIDNSHX=])/g;
while (file.readline(buf) >= 0) {
var m, t = buf.toString().split("\t");
if (t[0].charAt(0) == '@') continue;
if (chr_only && !/^(chr)?([0-9]+|X|Y)$/.test(t[2])) continue;
var flag = parseInt(t[1]);
if (flag&0x100) continue;
if (first_only && last_qname == t[0]) continue;
if (t[2] == '*') {
++n_unmapped;
continue;
} else {
++n_pri;
if (last_qname != t[0]) {
++n_mapped;
last_qname = t[0];
}
}
var pos = parseInt(t[3]) - 1, intron = [];
while ((m = re_cigar.exec(t[5])) != null) {
var len = parseInt(m[1]), op = m[2];
if (op == 'N') {
intron.push([pos, pos + len]);
pos += len;
} else if (op == 'M' || op == 'X' || op == '=' || op == 'D') pos += len;
}
if (intron.length == 0) {
++n_sgl;
continue;
}
n_splice += intron.length;
var chr = anno[t[2]];
if (chr != null) {
for (var i = 0; i < intron.length; ++i) {
var o = Interval.find_ovlp(chr, intron[i][0], intron[i][1]);
if (o.length > 0) {
var hit = false;
for (var j = 0; j < o.length; ++j) {
var st_diff = intron[i][0] - o[j][0];
var en_diff = intron[i][1] - o[j][1];
if (st_diff < 0) st_diff = -st_diff;
if (en_diff < 0) en_diff = -en_diff;
if (st_diff <= l_fuzzy && en_diff <= l_fuzzy)
++n_splice_hit, hit = true;
if (hit) break;
}
if (print_ovlp) {
var type = hit? 'C' : 'P';
if (hit && print_err_only) continue;
var x = '[';
for (var j = 0; j < o.length; ++j) {
if (j) x += ', ';
x += '(' + o[j][0] + "," + o[j][1] + ')';
}
x += ']';
print(type, t[0], i+1, t[2], intron[i][0], intron[i][1], x);
}
} else {
++n_splice_novel;
if (print_ovlp)
print('N', t[0], i+1, t[2], intron[i][0], intron[i][1]);
}
}
} else {
n_splice_novel += intron.length;
}
}
file.close();
buf.destroy();
if (!print_ovlp) {
print("# unmapped reads: " + n_unmapped);
print("# mapped reads: " + n_mapped);
print("# primary alignments: " + n_pri);
print("# singletons: " + n_sgl);
print("# predicted introns: " + n_splice);
print("# non-overlapping introns: " + n_splice_novel);
print("# correct introns: " + n_splice_hit + " (" + (n_splice_hit / n_splice * 100).toFixed(2) + "%)");
}
}
// evaluate overlap sensitivity
function paf_ov_eval(args)
{
var c, min_ovlp = 2000, min_frac = 0.95, min_mapq = 10;
while ((c = getopt(args, "q:l:f:")) != null) {
if (c == 'q') min_mapq = parseInt(getopt.arg);
else if (c == 'l') min_ovlp = parseInt(getopt.arg);
else if (c == 'f') min_frac = parseFloat(getopt.arg);
}
if (args.length - getopt.ind < 2) {
print("Usage: sort -k6,6 -k8,8n to-ref.paf | paftools.js ov-eval [options] - <ovlp.paf>");
print("Options:");
print(" -l INT min overlap length [2000]");
print(" -q INT min mapping quality [10]");
print(" -f FLOAT min fraction of mapped length [0.95]");
exit(1);
}
var buf = new Bytes();
var file = args[getopt.ind] == '-'? new File() : new File(args[getopt.ind]);
var a = [], h = {};
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
var is_pri = false;
if (parseInt(t[11]) < min_mapq) continue;
for (var i = 12; i < t.length; ++i)
if (t[i] == 'tp:A:P')
is_pri = true;
if (!is_pri) continue;
for (var i = 1; i <= 3; ++i)
t[i] = parseInt(t[i]);
for (var i = 6; i <= 8; ++i)
t[i] = parseInt(t[i]);
if (t[3] - t[2] < min_ovlp || t[8] - t[7] < min_ovlp || (t[3] - t[2]) / t[1] < min_frac)
continue;
var ctg = t[5], st = t[7], en = t[8];
while (a.length > 0) {
if (a[0][0] == ctg && a[0][2] > st)
break;
else a.shift();
}
for (var j = 0; j < a.length; ++j) {
if (a[j][3] == t[0]) continue;
var len = (en > a[j][2]? a[j][2] : en) - st;
if (len >= min_ovlp) {
var key = a[j][3] < t[0]? a[j][3] + "\t" + t[0] : t[0] + "\t" + a[j][3];
h[key] = len;
}
}
a.push([ctg, st, en, t[0]]);
}
file.close();
file = new File(args[getopt.ind + 1]);
while (file.readline(buf) >= 0) {
var t = buf.toString().split("\t");
var key = t[0] < t[5]? t[0] + "\t" + t[5] : t[5] + "\t" + t[0];
if (h[key] > 0) h[key] = -h[key];
}
file.close();
buf.destroy();
var n_ovlp = 0, n_missing = 0;
for (var key in h) {
++n_ovlp;
if (h[key] > 0) ++n_missing;
}
print(n_ovlp + " overlaps inferred from the reference mapping");
print(n_missing + " missed by the read overlapper");
print((100 * (1 - n_missing / n_ovlp)).toFixed(2) + "% sensitivity");
}
/*************************
***** main function *****
*************************/
function main(args)
{
if (args.length == 0) {
print("Usage: paftools.js <command> [arguments]");
print("Commands:");
print(" view convert PAF to BLAST-like (for eyeballing) or MAF");
print(" splice2bed convert spliced alignment in PAF/SAM to BED12");
print(" sam2paf convert SAM to PAF");
print(" delta2paf convert MUMmer's delta to PAF");
print(" gff2bed convert GTF/GFF3 to BED12");
print("");
print(" stat collect basic mapping information in PAF/SAM");
print(" asmstat collect basic assembly information");
print(" asmgene evaluate gene completeness (EXPERIMENTAL)");
print(" liftover simplistic liftOver");
print(" call call variants from asm-to-ref alignment with the cs tag");
print(" bedcov compute the number of bases covered");
print(" version print paftools.js version");
print("");
print(" mapeval evaluate mapping accuracy using mason2/PBSIM-simulated FASTQ");
print(" mason2fq convert mason2-simulated SAM to FASTQ");
print(" pbsim2fq convert PBSIM-simulated MAF to FASTQ");
print(" junceval evaluate splice junction consistency with known annotations");
print(" ov-eval evaluate read overlap sensitivity using read-to-ref mapping");
exit(1);
}
var cmd = args.shift();
if (cmd == 'view') paf_view(args);
else if (cmd == 'sam2paf') paf_sam2paf(args);
else if (cmd == 'delta2paf') paf_delta2paf(args);
else if (cmd == 'splice2bed') paf_splice2bed(args);
else if (cmd == 'gff2bed') paf_gff2bed(args);
else if (cmd == 'stat') paf_stat(args);
else if (cmd == 'asmstat') paf_asmstat(args);
else if (cmd == 'asmgene') paf_asmgene(args);
else if (cmd == 'liftover' || cmd == 'liftOver') paf_liftover(args);
else if (cmd == 'vcfpair') paf_vcfpair(args);
else if (cmd == 'call') paf_call(args);
else if (cmd == 'mapeval') paf_mapeval(args);
else if (cmd == 'bedcov') paf_bedcov(args);
else if (cmd == 'mason2fq') paf_mason2fq(args);
else if (cmd == 'pbsim2fq') paf_pbsim2fq(args);
else if (cmd == 'junceval') paf_junceval(args);
else if (cmd == 'ov-eval') paf_ov_eval(args);
else if (cmd == 'version') print(paftools_version);
else throw Error("unrecognized command: " + cmd);
}
main(arguments);