KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit Anti-GINS; Antigen: purified GINS complex | Sparks et al., 2019 | N/A |
| Rabbit Anti-Lrr1; Antigen: ACYQFLDKYLQSTRV | Dewar et al., 2017 | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| [α-32P]dATP | Perkin Elmer | Cat# BLU512H500UC |
| ATP | Sigma | Cat# A-5394 |
| Phosphocreatine | Sigma | Cat# P-6502 |
| Creatine Phosphokinase | Sigma | Cat# C-3755 |
| High Speed Supernatant (HSS) | Sparks and Walter, 2018 | N/A |
| Nucleoplasmic Extract (NPE) | Sparks and Walter, 2018 | N/A |
| Proteinase K | Roche | Cat# 3115879001 |
| RNase | Sigma | Cat# R4642-250mg |
| GINS expressed from pGC128 | Sparks et al., 2019 | N/A |
| Fen1mKikGR | Loveland et al., 2012 | N/A |
| LacI-biotin | Dewar et al., 2015 | N/A |
| p97-i (NMS873) | Sigma | Cat# SML 1128 |
| Cul-i (MLN4924) | Active Biochem | Cat# A-1139 |
| IPTG | Sigma | Cat# I5502 |
| Leupeptin | Roche | Cat# 11529048001 |
| Aprotinin | Roche | Cat# 11583794001 |
| BSA | Fisher | Cat# BP1600-100 |
| L-cysteine | Fisher | Cat# ICN10144601 |
| Cycloheximide | Sigma | Cat# C7698-5G |
| Cytochalasin B | Sigma | Cat# C6762-10MG |
| 14 mL Round-bottom falcon tubes | Fisher | Cat# 352059 |
| 2.5 mL thin-walled ultracentrifuge tubes | Beckman | Cat# 347356 |
| Nocodazole | Sigma | Cat# M1404-10MG |
| EDTA-free cOmplete protease inhibitor cocktail | Roche | Cat# 11873580001 |
| Nb.BbvCI | NEB | Cat# R0631S |
| Nt.BbvCI | NEB | Cat# R0632S |
| Nt.BspQI | NEB | Cat# R0644S |
| AflII | NEB | Cat# R0520S |
| NcoI | NEB | Cat# R0193L |
| MfeI-HF | NEB | Cat# R3589S |
| Klenow Fragment, exo- | NEB | Cat# M0212S |
| AlexaFluor647-maleimide | Thermo Fisher | Cat# A20347 |
| SYTOX Green | Thermo Fisher | Cat# S7020 |
| PD10 desalting column | GE Healthcare | Cat# 17-0851-01 |
| NiNTA resin | QIAGEN | Cat# 30410 |
| Streptavidin-coupled magnetic Dynabeads M-280 | Invitrogen | Cat# 11206D |
| Protein A Sepharose Fast Flow | GE Healthcare | Cat# 17127903 |
| Biotin-PEG-SVA | Laysan Bio | Cat# Bio-PEG-SVA-5k-100mg |
| M-PEG-SVA MW5000 | Laysan Bio | Cat# MPEG-SVA-5K-1g |
| Alt-R S.p. Cas9 H840A Nickase V3, 100 μg | IDT | Cat# 1081064 |
| Alt-R S.p. dCas9 Protein V3, 100 μg | IDT | Cat# 1081066 |
| Critical Commercial Assays | ||
| Bac-to-Bac Baculovirus Expression System | Thermo Fisher | Cat# 10359016 |
| MultiBac Expression System Kit | Geneva Biotech | Cat# MultiBac |
| ZR BAC DNA miniprep kit | Zymo Research | Cat# D4048 |
| Thermo Sequenase Cycle Sequencing Kits | USB | Cat# 78500 1KT |
| Experimental Models: Cell Lines | ||
| Sf9 Insect Cells | Expression Systems | Cat#94-001S |
| Tni Insect Cells | Expression Systems | Cat#94-002S |
| T7 Express Cells | NEB | Cat# C2566I |
| Experimental Models: Organisms/Strains | ||
| Xenopus laevis, adult female | Nasco | Cat# LM00535 |
| Xenopus laevis, adult male | Nasco | Cat# LM00715 |
| Oligonucleotides | ||
| Alt-R CRISPR-Cas9 crRNA XT (22 kb site, 5’-CAUGCCGUCACCCUAUUACA-3’) | This study (ordered from IDT) | N/A |
| Alt-R CRISPR-Cas9 crRNA XT (23 kb site, 5’-CAGGCGCUCCAUUGCCCAGU-3’) | This study (ordered from IDT) | N/A |
| Alt-R CRISPR-Cas9 crRNA XT (24 kb site, 5’-UGCCUGAGGCCAGUUUGCUC-3’) | This study (ordered from IDT) | N/A |
| Alt-R CRISPR-Cas9 crRNA XT (25 kb site, 5’-CCAGGUUCAACGGGCATGTA-3’) | This study (ordered from IDT) | N/A |
| Alt-R CRISPR-Cas9 tracrRNA, ATTO550 | This study (ordered from IDT) | Cat# 1075927 |
| Recombinant DNA | ||
| pKV44 | This study | N/A |
| pKV45 | This study | N/A |
| pGC261 | Sparks et al., 2019 | N/A |
| Software and Algorithms | ||
| NIS Elements | Nikon Instruments | https://www.nikoninstruments.com/Products/Software |
| MATLAB | Mathworks | https://www.mathworks.com/products/matlab.html |
| Bio-Formats | Linkert et al., 2010 | https://www.openmicroscopy.org/bio-formats/ |
| Image registration | Guiz-Sicairos et al., 2008 | https://doi.org/10.1364/OL.33.000156 |
| Molecule tracking script (u-track) | Jaqaman et al., 2008 | https://www.utsouthwestern.edu/labs/danuser/software/ |