Gibson Assembly Primer Designer (Beta)

Design primers with terminal overlaps for Gibson assembly of a vector and ordered DNA fragments.

Assembly primer inputs

Assembly setup

Includes the vector.

Editable overlap defaults depend on the total fragment count.

Vector

PCR opens after this base. Leave blank for an already-linear, blunt-ended vector.

Insert fragments

Enter fragments in assembly order, without added overlap tails.

Primer settings

Reaction settings

References

Kit guidance and primer Tm sources.

  1. Assembly guidance: NEB Gibson Assembly manual.
  2. Primer Tm: SantaLucia (1998); salt correction: Owczarzy et al. (2008).