// Copyright 2024 Google LLC // // Licensed under the Apache License, Version 2.0 (the "License"); // you may not use this file except in compliance with the License. // You may obtain a copy of the License at // // http://www.apache.org/licenses/LICENSE-2.0 // // Unless required by applicable law or agreed to in writing, software // distributed under the License is distributed on an "AS IS" BASIS, // WITHOUT WARRANTIES OR CONDITIONS OF ANY KIND, either express or implied. // See the License for the specific language governing permissions and // limitations under the License. syntax = "proto3"; package google.genomics.v1; import "google/api/annotations.proto"; import "google/genomics/v1/cigar.proto"; import "google/genomics/v1/position.proto"; import "google/protobuf/struct.proto"; option cc_enable_arenas = true; option go_package = "google.golang.org/genproto/googleapis/genomics/v1;genomics"; option java_multiple_files = true; option java_outer_classname = "ReadAlignmentProto"; option java_package = "com.google.genomics.v1"; // A linear alignment can be represented by one CIGAR string. Describes the // mapped position and local alignment of the read to the reference. message LinearAlignment { // The position of this alignment. Position position = 1; // The mapping quality of this alignment. Represents how likely // the read maps to this position as opposed to other locations. // // Specifically, this is -10 log10 Pr(mapping position is wrong), rounded to // the nearest integer. int32 mapping_quality = 2; // Represents the local alignment of this sequence (alignment matches, indels, // etc) against the reference. repeated CigarUnit cigar = 3; } // A read alignment describes a linear alignment of a string of DNA to a // [reference sequence][google.genomics.v1.Reference], in addition to metadata // about the fragment (the molecule of DNA sequenced) and the read (the bases // which were read by the sequencer). A read is equivalent to a line in a SAM // file. A read belongs to exactly one read group and exactly one // [read group set][google.genomics.v1.ReadGroupSet]. // // For more genomics resource definitions, see [Fundamentals of Google // Genomics](https://cloud.google.com/genomics/fundamentals-of-google-genomics) // // ### Reverse-stranded reads // // Mapped reads (reads having a non-null `alignment`) can be aligned to either // the forward or the reverse strand of their associated reference. Strandedness // of a mapped read is encoded by `alignment.position.reverseStrand`. // // If we consider the reference to be a forward-stranded coordinate space of // `[0, reference.length)` with `0` as the left-most position and // `reference.length` as the right-most position, reads are always aligned left // to right. That is, `alignment.position.position` always refers to the // left-most reference coordinate and `alignment.cigar` describes the alignment // of this read to the reference from left to right. All per-base fields such as // `alignedSequence` and `alignedQuality` share this same left-to-right // orientation; this is true of reads which are aligned to either strand. For // reverse-stranded reads, this means that `alignedSequence` is the reverse // complement of the bases that were originally reported by the sequencing // machine. // // ### Generating a reference-aligned sequence string // // When interacting with mapped reads, it's often useful to produce a string // representing the local alignment of the read to reference. The following // pseudocode demonstrates one way of doing this: // // out = "" // offset = 0 // for c in read.alignment.cigar { // switch c.operation { // case "ALIGNMENT_MATCH", "SEQUENCE_MATCH", "SEQUENCE_MISMATCH": // out += read.alignedSequence[offset:offset+c.operationLength] // offset += c.operationLength // break // case "CLIP_SOFT", "INSERT": // offset += c.operationLength // break // case "PAD": // out += repeat("*", c.operationLength) // break // case "DELETE": // out += repeat("-", c.operationLength) // break // case "SKIP": // out += repeat(" ", c.operationLength) // break // case "CLIP_HARD": // break // } // } // return out // // ### Converting to SAM's CIGAR string // // The following pseudocode generates a SAM CIGAR string from the // `cigar` field. Note that this is a lossy conversion // (`cigar.referenceSequence` is lost). // // cigarMap = { // "ALIGNMENT_MATCH": "M", // "INSERT": "I", // "DELETE": "D", // "SKIP": "N", // "CLIP_SOFT": "S", // "CLIP_HARD": "H", // "PAD": "P", // "SEQUENCE_MATCH": "=", // "SEQUENCE_MISMATCH": "X", // } // cigarStr = "" // for c in read.alignment.cigar { // cigarStr += c.operationLength + cigarMap[c.operation] // } // return cigarStr message Read { // The server-generated read ID, unique across all reads. This is different // from the `fragmentName`. string id = 1; // The ID of the read group this read belongs to. A read belongs to exactly // one read group. This is a server-generated ID which is distinct from SAM's // RG tag (for that value, see // [ReadGroup.name][google.genomics.v1.ReadGroup.name]). string read_group_id = 2; // The ID of the read group set this read belongs to. A read belongs to // exactly one read group set. string read_group_set_id = 3; // The fragment name. Equivalent to QNAME (query template name) in SAM. string fragment_name = 4; // The orientation and the distance between reads from the fragment are // consistent with the sequencing protocol (SAM flag 0x2). bool proper_placement = 5; // The fragment is a PCR or optical duplicate (SAM flag 0x400). bool duplicate_fragment = 6; // The observed length of the fragment, equivalent to TLEN in SAM. int32 fragment_length = 7; // The read number in sequencing. 0-based and less than numberReads. This // field replaces SAM flag 0x40 and 0x80. int32 read_number = 8; // The number of reads in the fragment (extension to SAM flag 0x1). int32 number_reads = 9; // Whether this read did not pass filters, such as platform or vendor quality // controls (SAM flag 0x200). bool failed_vendor_quality_checks = 10; // The linear alignment for this alignment record. This field is null for // unmapped reads. LinearAlignment alignment = 11; // Whether this alignment is secondary. Equivalent to SAM flag 0x100. // A secondary alignment represents an alternative to the primary alignment // for this read. Aligners may return secondary alignments if a read can map // ambiguously to multiple coordinates in the genome. By convention, each read // has one and only one alignment where both `secondaryAlignment` // and `supplementaryAlignment` are false. bool secondary_alignment = 12; // Whether this alignment is supplementary. Equivalent to SAM flag 0x800. // Supplementary alignments are used in the representation of a chimeric // alignment. In a chimeric alignment, a read is split into multiple // linear alignments that map to different reference contigs. The first // linear alignment in the read will be designated as the representative // alignment; the remaining linear alignments will be designated as // supplementary alignments. These alignments may have different mapping // quality scores. In each linear alignment in a chimeric alignment, the read // will be hard clipped. The `alignedSequence` and // `alignedQuality` fields in the alignment record will only // represent the bases for its respective linear alignment. bool supplementary_alignment = 13; // The bases of the read sequence contained in this alignment record, // **without CIGAR operations applied** (equivalent to SEQ in SAM). // `alignedSequence` and `alignedQuality` may be // shorter than the full read sequence and quality. This will occur if the // alignment is part of a chimeric alignment, or if the read was trimmed. When // this occurs, the CIGAR for this read will begin/end with a hard clip // operator that will indicate the length of the excised sequence. string aligned_sequence = 14; // The quality of the read sequence contained in this alignment record // (equivalent to QUAL in SAM). // `alignedSequence` and `alignedQuality` may be shorter than the full read // sequence and quality. This will occur if the alignment is part of a // chimeric alignment, or if the read was trimmed. When this occurs, the CIGAR // for this read will begin/end with a hard clip operator that will indicate // the length of the excised sequence. repeated int32 aligned_quality = 15; // The mapping of the primary alignment of the // `(readNumber+1)%numberReads` read in the fragment. It replaces // mate position and mate strand in SAM. Position next_mate_position = 16; // A map of additional read alignment information. This must be of the form // map (string key mapping to a list of string values). map info = 17; }