KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Monoclonal ANTI-FLAG M2-Peroxidase (HRP) antibody produced in mouse | Sigma-Aldrich | A8592–1MG; RRID:AB_439702 |
| Monoclonal Anti-HA antibody produced in mouse | Sigma-Aldrich | H3663–100UL; RRID:AB_262051 |
| Goat Anti-Mouse Goat anti-mouse IgG-HRP Polyclonal, Hrp Conjugated antibody | Santa Cruz Biotechnology | sc-2005; RRID:AB_631736 |
| PGK1 antibody [22C5D8] | abcam | ab113687; RRID:AB_10861977 |
| Monoclonal ANTI-FLAG M2 antibody produced in mouse | Sigma-Aldrich | F3165–1MG; RRID:AB_259529 |
| Monoclonal Anti-c-Myc antibody produced in mouse | Sigma-Aldrich | M4439–100UL; RRID:AB_439694 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Anhydrotetracycline hydrochloride (ahTET) | Acros Organics | AC23313–1000 |
| cOmplete, Mini Protease Inhibitor Cocktail | Roche | 4693124001 |
| Protein G-Agarose | Roche | 11243233001 |
| Ribonuclease A from bovine pancreas | Sigma-Aldrich | R6513–250MG |
| Easytides dATP, [α-32P] | PerkinElmer | BLU512H250UC |
| Hydroxyurea (HU) | Sigma-Aldrich | H8627–100G |
| Camptothecin (CPT) | Sigma-Aldrich | C9911–250MG |
| Phleomycin | InvivoGen | ant-ph-5 |
| Methyl methanesulfonate (MMS) | Sigma-Aldrich | M4016–1G |
| Doxycycline hyclate | Sigma-Aldrich | D9891–5G |
| SNAP-Surface Alexa Fluor 546 | NEB | S9129S |
| Chitin resin | NEB | S6651L |
| cOmplete His-tag resin | Roche | 5893682001 |
| SP Sepharose | GE Healthcare | 28950513 |
| anti-FLAG-M2 resin | Sigma | A1205 |
| PD10 desalting column | GE Healthcare | 17085101 |
| DOPC | Avanti Polar Lipids | 850375C |
| biotinylated-PE | Avanti Polar Lipids | 860562C |
| mPEG 2OOO-DOPE | Avanti Polar Lipids | 880130C |
| quartz slides | Finkenbeiner | N/A |
| Polymethylmethacrylate 25 kDa | Polymer Source | P9790-MMA |
| 3% PMMA 495 kDa | MICROCHEM | N/A |
| AquaSave | Mitsubishi Rayon Co., Ltd | N/A |
| λ-dsDNA | NEB | N3011S |
| M13mp18 ssDNA | NEB | N4040S |
| TLC plates | Millipore | HX71732079 |
| Critical Commercial Assays | ||
| amfiSure ONE PCR Master Mix (2×) | GenDEPOT | P7000–010 |
| In-Fusion HD Cloning Plus | TaKaRa Bio | 638909 |
| Ex Taq DNA Polymerase | TaKaRa Bio | RR001A |
| NucleoSpin Gel and PCR clean-up | MACHEREY-NAGEL | 740609.250 |
| RadPrime DNA Labeling System | Invitrogen | 18428–011 |
| PowerUp SYBR Green Master Mix | Applied Biosystems | A25742 |
| Amersham ECL Prime Western Blotting Detection Reagent | GE Healthcare | RPN2232 |
| Deposited Data | ||
| Unprocessed autoradiographs, western blot images and fluorescence microscopy images of this study | This study | https://doi.org/10.17632/g566xfgd96.1 |
| Experimental Models: Organisms/Strains | ||
| Fission and budding yeast strains used in this study are listed in Table S1 and Table S2. | N/A | N/A |
| Rosetta (DE3)pLys | Novagen | 70956–3 |
| yeast strain BJ5464 | Judith Campbell | N/A |
| SF9 insect cells | ThermoFisher | B82501 |
| DH10Bac | Invitrogen | N/A |
| Turbo E. coli | NEB | C2984H |
| BL21DE3 | Novagen | 69449–3 |
| Oligonucleotides | ||
| Oligonucleotides used in this study are listed in Table S1 | Sigma-Aldrich | N/A |
| Recombinant DNA | ||
| SNAP-34-Rad52 | Gibbet al., 2014 | Available upon request |
| pLK79 | Niu et al., 2010 | Available upon request |
| pRSF-Duet-Rmi1 | Niu et al., 2010 | Available upon request |
| pGAL10-DNA2 | Niu et al., 2010 | Available upon request |
| pFastBac-HTB-GFP-Sgs1 | Xue et al., 2019 | Available upon request |
| p11d-tscRPA-30MxeHis6 | Gibbet al., 2014 | Available upon request |
| pET21a-Exo1 | This study | Available upon request |
| pCM189-RAD52-HA | This study | Available upon request |
| pBluescriptKS (+) | Agilent | 212205 |
| pBluescriptKS(+)-arg1 -5’-λ1-I-PpolCS-λ2-hph-arg1-3’ | This study | Available upon request |
| Software and Algorithms | ||
| Image Quant TL 7.0 | GE Healthcare | N/A |
| Prism 6 | GraphPad | N/A |
| Prism 8 | GraphPad | N/A |
| NIS-Element software | Nikon | N/A |
| FIJI | ImageJ software | N/A |
| Other | ||
| Edinburgh Minimal Media Glutamate (EMMG) (Powder) | United States Biological | E2205–15 |
| Storage Phosphor Screen | Molecular Dynamics | N/A |
| Typhoon TRIO Variable Mode Imager | GE Healthcare | N/A |
| MiniBeadBeater-16, Model 607 | BioSpec Products | N/A |
| Misonix Sonicator 3000 Ultrasonic Cell Disruptors | Misonix | N/A |
| 7900HT Fast Real-Time PCR system | Applied Biosystems | N/A |
| MicroAmp Fast Optical 96-Well Reaction Plate, 0.1 mL | Applied Biosystems | 4346907 |
| BX51 microscope | Olympus | N/A |
| AxioCam MRm camera | Zeiss | N/A |
| Azure c400 Imaging System | Azure Biosystems | N/A |
| Inverted TIRF Microscope | Nikon | N/A |
| 488 nm laser | Coherent Sapphire | N/A |
| 561 nm laser | Coherent Sapphire | N/A |
| iXon X3 EMCCD cameras | Andor | N/A |
| GeneScreen Plus Hybridization Transfer Membrane | PerkinElmer | NEF988001PK |
| Westran S, 0.2 μm PVDF membrane | Whatman | 10413096 |