If you work with plant transformation, particularly in Arabidopsis thaliana or related species, you’ve likely encountered the PGPS3 vector. Derived from the pGreen series, PGPS3 is a compact, high-copy-number binary vector known for its small size (approximately 3.2 kb), making it ideal for golden gate cloning, site-directed mutagenesis, and efficient transformation into Agrobacterium tumefaciens .
| Restriction Enzyme | Position (approx.) | Recognition Sequence | Best Use Case | |-------------------|-------------------|----------------------|----------------| | | 1,234 | G^AATTC | Insert cloning (5' end) | | BamHI | 1,567 | G^GATCC | Insert cloning (3' end) | | HindIII | 2,345 | A^AGCTT | Backbone linearization | | XbaI | 789 | T^CTAGA | Subcloning from other vectors | | SacI | 2,890 | GAGCT^C | Terminator swaps | | PstI | 3,101 | CTGCA^G | Rare cutter for large inserts | | KpnI | 456 | GGTAC^C | Directional cloning with SacI | | NotI | 2,567 | GC^GGCCGC | For GC-rich inserts (rare cutter) | pgps3 unique restriction sites
Happy cloning! Have you found a rare unique site in PGPS3 that I missed? Or a tricky methylation issue? Drop a comment below or tag me on Twitter @PlantBiotechLab – let’s build a better restriction map together. If you work with plant transformation, particularly in
However, successful cloning depends entirely on one thing: knowing which restriction enzymes cut once —and only once—in your plasmid. Have you found a rare unique site in PGPS3 that I missed
Just remember: no SalI, no EcoRV, and always verify your map. When in doubt, a quick virtual digest takes 30 seconds and saves three days of failed ligations.
| Enzyme | Problem | |--------|---------| | | Cuts twice (once in MCS, once in the pSa origin of replication). | | EcoRV | Cuts three times (unexpectedly in the LacZ alpha fragment and the kanamycin resistance gene). | | ClaI | Cuts twice (in the MCS and the ColE1 origin). |