Differences between pUPD and pUPD2
The table below summarizes the differences between pUPD and pUPD2:
| pUPD | pUPD2 | |
|---|---|---|
| Backbone | pGEMT | pSB1C3 |
| Resistance | Ampicillin | Chloramphenicol |
| 5' clooning site | CTCG | CTCG |
| 3' cloning site | CGAG | TGAG |
| Sequencing primers | T7, SP6 | No universal primers |
Important things to take into account when moving to pUPD2:
1. The resistance has changed from ampicillin to chloramphenicol.
2. The pUPD2 does not have any universal primer binding site for sequencing. Our sequencing primers are: 5'-GCTTTCGCTAAGGATGATTTCTGG-3' (forward) and 5'-CAGGGTGGTGACACCTTGCC-3' (reverse).
3. IMPORTANT: GB2.0 and GB3.0 parts are fully compatible. In other words, GBparts domesticated in the old pUPD vector can be combined with new parts assembled in the pUPD2 vector to create new TUs.
4. However, PCR products designed with GB2.0 version of the GB Domesticator tool (intended to be cloned into old pUPD version) cannot be cloned into pUPD2 because of a change in the 3' cloning site. This modification was introduced to prevent the entry of PCR fragments in reverse orientation.
