Table 1.
Expression plasmids used in this protocol
| Plasmid # | Description | Vector backbone |
Antibiotic marker |
|---|---|---|---|
| pAM81 [32] | Rpnl, Rpn2, Rpn13 | pETDuet | Ampicillin |
| pAM82 [32] | FLAG-Rpt1, Rpt2, 6xHis-Rpt3, Rpt5, Rpt6, Rpt4 | pCOLADuet Kanamycin | |
| pAM83 [32] | RIL rare tRNAs, Nas6, Nas2, Hsm3, Rpn14 | pACYCDuet | Chloramphenicol |
| pAM85 (modified from [33]) | Rpn9, Rpnll, Rpn8, MBP-HRV-Rpn6, Rpn5 | pETDuet | Ampicillin |
| pAM86 (modified from[33]) | 6xHis-HRV-Rpn12, Rpn7, Rpn3 | pCOLADuet Kanamycin | |
| pAM80 [33] | Seml, Hsp90 | pACYCDuet | Chloramphenicol |
| pAM87 (constructed from [24, 25]) | pAzFRS.2.tl, UAG tRNA | pCDF | Spectinomycin |
| pAM82 (modified from [32]) | pAM82 with the UAG codon inserted at the desired location | pCOLADuet | Kanamycin |