Add or pick modules, choose an assembly route, then domesticate sites and assign seams to get a complete circular plasmid with primers and risk notes.
From gene modules to a complete plasmid, five stages annotate each step in this entry.
Build a module from a host, promoter, CDS, and terminator — or paste a whole sequence.
Multi-select, reorder, and set each module's direction.
Pick a backbone, a selection marker, and the assembly route (auto, in-vitro, or in-vivo).
Type IIS sites (BsaI/BsmBI/BbsI) are scanned and cleaned automatically.
Seams are assigned, the circular plasmid is assembled, and primers are designed.
In-vitro Golden Gate, hierarchical MoClo, Gibson, and in-vivo recombination; auto selects by module count and host.
Type IIS sites (BsaI/BsmBI/BbsI) are scanned and cleaned so they do not block the assembly.
Seams are assigned between modules; colony-PCR and sequencing primers are generated.
GC windows, repeats, and stem-loop structures are flagged before you order.
Add modules in the area below, then select, order, and assemble them.
Watch the assembly advance stage by stage.
The assembled plasmid with route, seams, domestication report, colony-PCR primers, and risk list.
Each saved run becomes a reproducible entry with an Entry ID you can cite in your methods. This tool saves the assembled plasmid — no downstream hand-off.
Run an assembly first, then save it here.