Enzymes & cloning

Golden Gate from part plasmids

Golden Gate as the reaction runs: digest pre-domesticated part plasmids with a Type IIS enzyme and assemble them in the order their OVERHANGS dictate. The fragment released from each part is the one carrying no recognition site (the site goes out with the backbone, which is why a mis-ordered assembly is not re-cut), and the assembly order is an OUTPUT — a set whose overhangs do not close into a single cycle has no product, and the reason is the answer. Distinct from cloning_simulate's goldengate method, which does the other job: designing the primers that ADD the sites to BARE parts, assembled in the order you list them.

post/golden_gate_from_parts

Request

  • Base URL: https://seqbench.com/api/v1
  • URL: https://seqbench.com/api/v1/golden_gate_from_parts
  • Auth: none required

Request body

partsobject[] required

The part plasmids: [{ name, sequence, circular }]. Each needs two Type IIS sites facing INWARDS at the part. circular defaults to true, which is what a level-0 part plasmid is. 2-24 parts, each at most 30,000 bp and 200,000 bp in total.

enzyme'BsaI' | 'BsmBI' | 'BbsI' | 'SapI'

Type IIS enzyme (BsaI, BsmBI, BbsI, SapI). NEB's variant names (BsaI-HFv2, BbsI-HF, BsmBI) fold to the parent enzyme; any other name is rejected rather than substituted, since site, spacer and overhang length are all enzyme-specific.

Response

Tool ran successfully.

okunknown required
toolstring required

The tool slug that ran.

resultobject required

Tool-specific output object. Its shape depends on the tool — call GET /{tool} or see the docs for each tool's fields.

Example response

{
  "provenance": {
    "apiVersion": "1.1.0"
  }
}

Changes

No recorded changes to this operation across all 1 revision of this API.