/***************************************************************** * The K8 Javascript interpreter is required to run this script. * * * * Source code: https://github.com/attractivechaos/k8 * * Binary: http://sourceforge.net/projects/lh3/files/k8/ * * * * Data file used for generating GRCh38 ALT alignments: * * * * http://sourceforge.net/projects/bio-bwa/files/ * *****************************************************************/ /****************** *** From k8.js *** ******************/ 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; } /************************ *** command markovlp *** ************************/ function bwa_markOvlp(args) { var c, min_aln_ratio = .9, min_ext = 50; while ((c = getopt(args, "r:e:")) != null) { if (c == 'r') min_aln_ratio = parseFloat(getopt.arg); else if (c == 'e') min_ext = parseInt(getopt.arg); } var file = args.length == getopt.ind? new File() : new File(args[getopt.ind]); var buf = new Bytes(); var dir4 = ['>>', '><', '<>', '<<']; while (file.readline(buf) >= 0) { var t = buf.toString().split("\t") for (var i = 0; i < t.length; ++i) if (i != 0 && i != 4) t[i] = parseInt(t[i]); var el, a1, a2, e1, e2, r1, r2; // a: aligned length; e: extended length; r: remaining length e2 = a2 = t[7] - t[6]; if (t[2] < t[3]) { // forward-forward match e1 = a1 = t[3] - t[2]; r1 = t[2] - t[6]; r2 = (t[5] - t[7]) - (t[1] - t[3]); el = r1 > 0? t[6] : t[2]; el += r2 > 0? t[1] - t[3] : t[5] - t[7]; } else { // reverse-forward match e1 = a1 = t[2] - t[3]; r1 = (t[1] - t[2]) - t[6]; r2 = (t[5] - t[7]) - t[3]; el = r1 > 0? t[6] : t[1] - t[2]; el += r2 > 0? t[3] : t[5] - t[7]; } e1 += el; e2 += el; var type; if (a1 / e1 >= min_aln_ratio && a2 / e2 >= min_aln_ratio) { if ((r1 >= min_ext && r2 >= min_ext) || (r1 <= -min_ext && r2 <= -min_ext)) { // suffix-prefix match var d = t[2] < t[3]? 0 : 2; if (r1 < 0) d ^= 3; // reverse the direction type = 'O' + dir4[d]; } else type = 'C' + (e1 / t[1] > e2 / t[5]? 1 : 2); } else type = 'I'; // internal local match; not a suffix-prefix match //print(t[1], e1, a1, t[5], e2, a2); print(type, buf); } buf.destroy(); file.close(); } /*********************** *** command pas2bed *** ***********************/ function bwa_pas2reg(args) { var file = args.length? new File(args[0]) : new File(); var buf = new Bytes(); while (file.readline(buf) >= 0) { var t = buf.toString().split("\t"); if (t[0] == t[4]) continue; if (parseInt(t[2]) < parseInt(t[3])) print(t[0], t[1], t[2], t[3], t[8]); else print(t[0], t[1], t[3], t[2], t[8]); print(t[4], t[5], t[6], t[7], t[8]); } buf.destroy(); file.close(); } /******************* * command sam2pas * *******************/ function bwa_sam2pas(args) { var file = args.length == 0? new File() : new File(args[0]); var buf = new Bytes(); var seq_dict = {}; while (file.readline(buf) >= 0) { var line = buf.toString(); var m; if (/^@SQ/.test(line)) { var name = null, len = null; if ((m = /\tSN:(\S+)/.exec(line)) != null) name = m[1]; if ((m = /\tLN:(\S+)/.exec(line)) != null) len = parseInt(m[1]); if (name != null && len != null) seq_dict[name] = len; } if (/^@/.test(line)) continue; var t = line.split("\t"); var pos = parseInt(t[3]) - 1; var x = 0, y = 0, i = 0, clip = [0, 0], n_ins = 0, n_del = 0, o_ins = 0, o_del = 0, n_M = 0; var re = /(\d+)([MIDSH])/g; while ((m = re.exec(t[5])) != null) { var l = parseInt(m[1]); if (m[2] == 'M') x += l, y += l, n_M += l; else if (m[2] == 'I') y += l, n_ins += l, ++o_ins; else if (m[2] == 'D') x += l, n_del += l, ++o_del; else if (m[2] == 'S' || m[2] == 'H') clip[i == 0? 0 : 1] = l; ++i; } var is_rev = (parseInt(t[1]) & 16)? true : false; var misc = 'mapQ=' + t[4] + ';'; var usc = 1; if ((m = /\tNM:i:(\d+)/.exec(line)) != null) { var NM = parseInt(m[1]); var diff = (NM / (n_M + n_ins + n_del)).toFixed(3); misc += 'diff=' + diff + ';n_mis=' + (NM - n_del - n_ins) + ';'; } misc += 'n_del='+n_del+';n_ins='+n_ins+';o_del='+o_del+';o_ins='+o_ins + ';'; if ((m = /\tAS:i:(\d+)/.exec(line)) != null) { misc += 'AS='+m[1] + ';'; usc = (parseInt(m[1]) / (x > y? x : y)).toFixed(3); } if ((m = /\tXS:i:(\d+)/.exec(line)) != null) misc += 'XS='+m[1] + ';'; var len = y + clip[0] + clip[1]; var z = [t[0], len, clip[0], clip[0] + y, t[2], seq_dict[t[2]], pos, pos + x, usc, misc]; if (parseInt(t[1]) & 16) z[2] = clip[1] + y, z[3] = clip[1]; print(z.join("\t")); } buf.destroy(); file.close(); } /*********************** *** command reg2cut *** ***********************/ function bwa_reg2cut(args) { var c, min_usc = 0.5, min_ext = 100, min_len = 5000, cut = 250; while ((c = getopt(args, "s:e:l:c:")) != null) { if (c == 's') min_usc = parseFloat(getopt.arg); else if (c == 'e') min_ext = parseInt(getopt.arg); else if (c == 'l') min_len = parseInt(getopt.arg); else if (c == 'c') cut = parseInt(getopt.arg); } var file = args.length == getopt.ind? new File () : new File(args[getopt.ind]); var buf = new Bytes(); function print_bed() { for (var i = 0; i < a.length; ++i) { var start = a[i][0] - cut > 0? a[i][0] : 0; var end = a[i][1] + cut < last_len? a[i][1] : last_len; if (end - start >= min_len) print(last_chr, start, end); } } var last_chr = null, last_len = null, max_c_usc = 0, start = 0, end = 0; var a = []; while (file.readline(buf) >= 0) { var t = buf.toString().split("\t"); t[1] = parseInt(t[1]); t[2] = parseInt(t[2]); t[3] = parseInt(t[3]); t[4] = parseFloat(t[4]); var is_contained = t[2] < min_ext && t[1] - t[3] < min_ext? true : false; if (t[3] - t[2] < cut<<1) continue; t[2] += cut; t[3] -= cut; if (t[0] != last_chr) { a.push([start, end]); if (last_chr != null && max_c_usc < min_usc) print_bed(); last_chr = t[0]; last_len = t[1]; start = t[2]; end = t[3]; max_c_usc = is_contained? t[4] : 0; a.length = 0; } else { if (is_contained) max_c_usc = max_c_usc > t[4]? max_c_usc : t[4]; if (t[4] < min_usc) continue; if (t[2] > end) { a.push([start, end]); start = t[2]; end = end > t[3]? end : t[3]; } else end = end > t[3]? end : t[3]; } } a.push([start, end]); if (max_c_usc < min_usc) print_bed(); // the last sequence buf.destroy(); file.close(); } function bwa_shortname(args) { var file = args.length? new File(args[0]) : new File(); var buf = new Bytes(); var re = /(\S+)\/(\d+)_(\d+)((:\d+-\d+)+)/g; var re2 = /:(\d+)-(\d+)/g; while (file.readline(buf) >= 0) { var match, match2; var line = buf.toString(); var x = []; while ((match = re.exec(line)) != null) { var start = parseInt(match[2]), len = parseInt(match[3]) - start; while ((match2 = re2.exec(match[4])) != null) { var a = parseInt(match2[1]) - 1; var b = parseInt(match2[2]); start += a; len = b - a; } x.push([match[0], match[1] + '/' + start.toString() + '_' + (start+len).toString()]); } for (var i = 0; i < x.length; ++i) line = line.replace(x[i][0], x[i][1]); print(line); } buf.destroy(); file.close(); } /******************* * Command gff2sam * *******************/ function bwa_gff2sam(args) { if (args.length < 2) { print("Usage: k8 bwa-helper.js "); exit(1); } var file = new File(args[1]); var buf = new Bytes(); var len = {}; while (file.readline(buf) >= 0) { var t = buf.toString().split(/\s+/); len[t[0]] = parseInt(t[1]); } file.close(); file = new File(args[0]); var re_cigar = /([MID])(\d+)/g; var lineno = 0; while (file.readline(buf) >= 0) { ++lineno; var t = buf.toString().split("\t"); var m = /Target=(\S+)\s+(\d+)\s+(\d+)\s+([+-])/.exec(t[8]); if (m == null) { warn("WARNING: skipped line "+lineno+" due to the lack of Target."); continue; } var qname = m[1]; var flag = t[6] == m[4]? 0 : 16; var qb = parseInt(m[2]) - 1, qe = parseInt(m[3]), qlen = len[qname]; if (qlen == null) throw Error("Sequence "+qname+" is not present in the query-length.txt"); var clip5 = qb, clip3 = qlen - qe; if (flag&16) clip5 ^= clip3, clip3 ^= clip5, clip5 ^= clip3; // swap m = /Gap\s*=\s*(([MID]\d+\s*)+)/.exec(t[8]); var cigar = clip5? clip5 + 'S' : ''; var n_ins = 0, n_del = 0, n_match = 0, NM = null; if (m) { var mc; while ((mc = re_cigar.exec(m[1])) != null) { var l = parseInt(mc[2]); cigar += mc[2] + mc[1]; if (mc[1] == 'I') n_ins += l; else if (mc[1] == 'D') n_del += l; else if (mc[1] == 'M') n_match += l; } if (n_ins + n_match != qe - qb || n_del + n_match != parseInt(t[4]) - parseInt(t[3]) + 1) throw Error("Inconsistent CIGAR at line "+lineno); } else { // ungapped alignment var tb = parseInt(t[3]) - 1, te = parseInt(t[4]); if (te - tb != qe - qb) { warn("WARNING: line "+lineno+" should contain gaps, but lacks Gap. Skipped.\n"); } else cigar = (qe - qb) + 'M'; } if (clip3) cigar += clip3 + 'S'; if ((m = /num_mismatch=(\d+)/.exec(t[8])) != null) NM = parseInt(m[1]) + n_ins + n_del; var out = [qname, flag, t[0], t[3], 255, cigar, '*', 0, 0, '*', '*']; if (NM != null) out.push('NM:i:' + NM); print(out.join("\t")); } buf.destroy(); file.close(); } /********************* *** Main function *** *********************/ function main(args) { if (args.length == 0) { print("\nUsage: k8 bwa-helper.js [arguments]\n"); print("Commands: sam2pas convert SAM to pairwise alignment summary format (PAS)"); print(" pas2reg extract covered regions"); print(" reg2cut regions to extract for the 2nd round bwa-mem"); print(" markovlp identify bi-directional overlaps"); print(" gff2sam convert GFF3 alignment to SAM"); print(" shortname shorten sequence name after subseq (PacBio read names only)"); print(""); exit(1); } var cmd = args.shift(); if (cmd == 'sam2pas') bwa_sam2pas(args); else if (cmd == 'gff2sam') bwa_gff2sam(args); else if (cmd == 'markovlp') bwa_markOvlp(args); else if (cmd == 'pas2reg') bwa_pas2reg(args); else if (cmd == 'reg2cut') bwa_reg2cut(args); else if (cmd == 'shortname') bwa_shortname(args); else warn("Unrecognized command"); } main(arguments);