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Variant to construct to genotyping bridge

variant_to_construct
Read-onlyIdempotent

Turn one variant into one buildable plan: verify the reference allele actually sits where the coordinate says, apply the edit, design site-directed mutagenesis primers to install it, design KASP/ARMS allele-specific primers to genotype it afterwards, and consolidate everything into a single oligo order table. Takes either a construct sequence with a 1-based position and ref/alt alleles (offline, deterministic), or an HGVS "c." description resolved through the MANE crosswalk and a live Ensembl exon map. A mismatched reference allele is refused with the bases that were actually found there, because a coordinate that is right for another isoform yields a perfectly valid primer set for the wrong base. Bases shared by both alleles are trimmed first, so a VCF-anchored pair is designed as the substitution or indel it actually is. Mutagenesis covers every class (a substitution, an insertion, a deletion and a multi-base replacement are all one interval replacement); KASP needs a single-base substitution's 3'-terminal base, so for an indel the genotyping half comes back as a named omission with the reason and the readout that does work, never as an empty list.

Input Schema

TableJSON Schema
NameRequiredDescriptionDefault
flankNoNETWORK MODE. Bases of genomic context fetched either side of the variant (60-1000). Needs to cover the allele-specific core upstream and the genotyping amplicon downstream.
labelNoName stem for the ordered oligos. Defaults to the HGVS string (network mode) or a positional label like "A100G" (offline).
styleNoMutagenic primer style: overlapping (QuikChange) or back-to-back (Q5/KLD).quikchange
variantNoNETWORK MODE. A full HGVS "c." description, e.g. "NM_000546.6:c.215C>G" or "TP53:c.215C>G". Resolved through the bundled MANE crosswalk and a live Ensembl exon map, then a genomic window is fetched around it. Mutually exclusive with "sequence".
positionNoOFFLINE MODE. 1-based position of the first base of refAllele on `sequence`. For a pure insertion (empty refAllele) the alternate bases are inserted immediately BEFORE this position.
sequenceNoOFFLINE MODE. The reference/construct sequence the variant is described against (raw or FASTA). Mutually exclusive with "variant".
altAlleleNoOFFLINE MODE. Bases installed. Use "-" or "" for a pure deletion.
refAlleleNoOFFLINE MODE. Bases replaced, as they appear on `sequence` — checked against it and refused if they differ. Use "-" or "" for a pure insertion. A VCF-style anchored pair (both alleles carrying a shared flanking base, e.g. refAllele "T" / altAllele "TGGATCC", or "TG" / "AG") is accepted and trimmed to its minimal form before anything is designed; the trimmed coordinates are reported as `minimal`.
armTmTargetNoTarget Tm (°C) for each mutagenic primer's template-binding arm (45-80; the arm is grown between 10 and 30 nt, so a target outside that range is refused rather than clamped to the shortest or longest arm).
maxAmpliconNoMaximum genotyping amplicon length. Refused below 36 bp, the shortest KASP product that can exist (an 18 nt core plus an 18 nt reverse primer), and refused below minAmplicon — both are unsatisfiable for every reference, and left unchecked they came back as "no common reverse primer could be placed; provide more 3' flanking sequence".
minAmpliconNoMinimum genotyping amplicon length, measured from the allele-specific primer's 5' end (which sits one core-length upstream of the variant) to the reverse primer's 5' end. Must not exceed maxAmplicon.
targetCoreTmNoTarget Tm (°C) for the allele-specific genotyping core, before the universal tail (45-80; the core is chosen between 18 and 27 nt, so a target outside that range is refused rather than clamped).
addSecondaryMismatchNoEngineer the ARMS destabilising mismatch 3 nt from the allele-specific primer's 3' end.

TDQS

A4.5/5.0
Behavior5/5

Does the description disclose side effects, auth requirements, rate limits, or destructive behavior?

Far beyond the readOnly/idempotent/openWorld annotations, the description discloses that a mismatched reference allele is refused with the bases actually found, that shared allele bases are trimmed to minimal form, that mutagenesis treats all variant classes as one interval replacement, and that indel genotyping returns a named omission with the reason — never an empty list. These behaviors are also consistent with readOnlyHint=true since the tool produces a plan rather than mutating state.

Agents need to know what a tool does to the world before calling it. Descriptions should go beyond structured annotations to explain consequences.

Conciseness4/5

Is the description appropriately sized, front-loaded, and free of redundancy?

The description is dense but every sentence carries operational content: mode selection, isoform-coordinate risk, allele trimming, primer-class coverage, and output refusal behavior. It earns its length for a 13-parameter, two-mode tool, though presenting it as one continuous paragraph slightly reduces scannability compared to a structured breakdown.

Shorter descriptions cost fewer tokens and are easier for agents to parse. Every sentence should earn its place.

Completeness4/5

Given the tool's complexity, does the description cover enough for an agent to succeed on first attempt?

With no output schema, the description carries the burden of defining return behavior and largely succeeds: a consolidated oligo order table, minimal trimmed coordinates, refusal with the bases actually found, and a named omission with a working readout for indels. Remaining gaps — the exact field structure of the order table and network-mode failure behavior — are minor given that the safety profile is already covered by annotations.

Complex tools with many parameters or behaviors need more documentation. Simple tools need less. This dimension scales expectations accordingly.

Parameters4/5

Does the description clarify parameter syntax, constraints, interactions, or defaults beyond what the schema provides?

Schema coverage is 100% and each parameter description is already rich (bounds, refusal conditions, mode tagging, even a quoted error message). The main description adds the cross-parameter semantics the schema cannot express alone: that KASP requires a single-base substitution's 3'-terminal base, linking the ref/alt inputs to the shape of the genotyping output, and tying the trimming behavior to how anchored VCF pairs are interpreted.

Input schemas describe structure but not intent. Descriptions should explain non-obvious parameter relationships and valid value ranges.

Purpose5/5

Does the description clearly state what the tool does and how it differs from similar tools?

The description opens with a specific verb+resource: 'Turn one variant into one buildable plan' and enumerates the complete pipeline — verifying the reference allele, applying the edit, designing site-directed mutagenesis primers, designing KASP/ARMS genotyping primers, and consolidating into a single oligo order table. This clearly distinguishes it from sibling tools like site_directed_mutagenesis or kasp_primer_design, which cover only one stage of that pipeline.

Agents choose between tools based on descriptions. A clear purpose with a specific verb and resource helps agents select the right tool.

Usage Guidelines4/5

Does the description explain when to use this tool, when not to, or what alternatives exist?

The description clearly delimits the two mutually exclusive input modes (offline sequence+position vs network HGVS resolved through MANE/Ensembl) and the refusal/omission conditions that govern when results come back. It does not explicitly name sibling alternatives or state 'use X instead when you only need one stage,' but the consolidated 'bridge' purpose makes the scope reasonably inferable from context.

Agents often have multiple tools that could apply. Explicit usage guidance like "use X instead of Y when Z" prevents misuse.

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TDQS

A3.6/5.0
Disambiguation4/5

Most tools have highly specific purposes (e.g., crispr_grna_design vs base_editing_design vs prime_editing_design). However, there is some overlap in sequence analysis tools (characterize_sequence, sequence_report) and plasmid annotation tools (plasmid_annotate vs plasmid_deep_annotate) which could cause confusion.

Naming Consistency3/5

The naming pattern is largely consistent with snake_case verb_noun or noun_descriptor (e.g., primer_design, plasmid_annotate, fastq_trim). However, there are exceptions like 'batch', 'workflow', 'gc_content', and 'cloning_diagnose' which don't follow the verb_noun pattern consistently. Also, some names are phrases like 'golden_gate_from_parts'.

Tool Count2/5

With 101 tools, this server is extremely large and likely overwhelming for agents. Even for a comprehensive bioinformatics toolkit, this exceeds a manageable scope, risking agent confusion and inefficient tool selection. A more modular approach would be advisable.

Completeness4/5

The tool surface covers a wide range of bioinformatics workflows including sequence analysis, primer design, cloning, CRISPR, NGS, expression analysis, and data export. There are minor gaps such as lack of a dedicated protein structure prediction tool and limited off-target genome coverage, but overall the set is impressively complete for its domain.

Resources