KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| anti-FANCJ antibody | Novus Biologicals | Cat# NBP1-31883 |
| anti-Actin antibody | Santa Cruz | Cat# sc-47778; RRID: AB_626632 |
| anti-Flag antibody | Sigma | Cat# F3165-.2MG; RRID: AB_259529 |
| anti-xlFANCJ-N | (ref.35) | Pocono Rabbit Farm and Laboratory Custom Projects 28331 and 28332 |
| anti-xlFANCJ-C | This paper | Bethyl Laboratories Custom Project 61565A |
| anti-RTEL1-N | J. Walter laboratory, (ref. 14) | Pocono Rabbit Farm and Laboratory Custom Project Pocono 32259 |
| anti-CDC45 | J. Walter laboratory, (ref. 36) | Pocono Rabbit Farm and Laboratory Custom Project 534 |
| anti-M.HpaII | J. Walter laboratory, (ref. 6) | Pocono Rabbit Farm and Laboratory Custom Project 31495 |
| anti-PSMA3 | J. Walter laboratory, (ref. 6) | New England Peptide Custom Project 3516 |
| anti-SPRTN-N | J. Walter laboratory, (ref. 6) | Pocono Rabbit Farm and Laboratory Custom Project 31053 |
| anti-Histone H3 | Cell Signaling | Cat# 9715S; RRID: AB_331563 |
| anti-Mcm6 | J. Walter laboratory, (ref. 14) | New England Peptide Custom Project 2926 |
| anti-HMCES | J.Walter laboratory, (ref. 40) | New England peptide Custom Project 4377 |
| Bacterial and virus strains | ||
| BL21 (DE3) | Thermo Scientific | Cat# C600003 |
| Rosetta (DE3) Escherichia coli | MilliporeSigma™ | Cat# 70-954-3 |
| Chemicals, peptides, and recombinant proteins | ||
| Triton X-100 | Sigma | Cat# T8787-250ML |
| 16% Formaldehyde (w/v), Methanol-free | Thermo Scientific | Cat# 28906 |
| 4x NuPAGE LDS sample buffer | Thermo Scientific | Cat# NP0007 |
| smDNAse | Max-Planck-Institute for Biochemistry | N/A |
| cOmplete EDTA-free protease inhibitor cocktail | Sigma | Cat# 4693132001 |
| DTT | Roth | Cat# 6908.2 |
| His-TEV protease | This paper | N/A |
| Pefabloc SC | Sigma | Cat# 76307-1G |
| Biotin | IBA Lifesciences | Cat# 2-1016-005 |
| isopropyl-β-D-thiogalactoside | Sigma | Cat# I6758-10G |
| Imidazole | Roth | Cat# 3899.1 |
| IGEPAL | Sigma | Cat# I8896-50ML |
| Protino Ni-NTA Agarose | Fisher Scientific | Cat# 11912422 |
| Tween20 | Sigma | Cat# P7949 |
| NaSCN | Sigma | Cat# 467871 |
| Q5® High-Fidelity DNA Polymerase | New England BioLabs | Cat# M0515 |
| UltraPure BSA | Thermo Scientific | Cat# AM2616 |
| UDG | New England BioLabs | Cat# M0280L |
| Orange G | Sigma | Cat# O7252-25G |
| Protein G | BioVision | Cat# 6510 |
| ATP | Thermo Fischer | Cat# R0441 |
| Trypsin Gold | Promega | Cat# V5280 |
| HaeIII | New England BioLabs | Cat# R0108S |
| dNTPs | New England BioLabs | Cat# N0447S |
| UREA | Roth | Cat# 3941.3 |
| Ficoll | Sigma | Cat# F4375-25G |
| Lipofectamine 2000 | Invitrogen | Cat# 11668019 |
| Puromycin | Gibco | Cat# A1113803 |
| Q5 Master Mix | New England BioLabs | Cat# M0494L |
| BP clonase | Thermo Fischer | Cat# 11789020 |
| LR clonase | Thermo Fischer | Cat# 11791100 |
| Hygromycin B | Fisher Scientific | Cat# 10687010 |
| Doxycycline Hyclate | Sigma | Cat# D9891 |
| TCEP | Roth | Cat# HN95.3 |
| HRV 3C protease | Thermo Fisher | Cat# 88947 |
| 3XFlag peptide | Sigma | Cat# F4799-4MG |
| ESF 921 insect cell culture medium | Fisher scientific | Cat# 96-001-01-CS |
| Nt.BbvcI | New England BioLabs | Cat# R0632L |
| M.HpaII | J. Walter laboratory, (ref. 5) | N/A |
| Exonuclease I | New England BioLabs | Cat# M0293S |
| S-adenosylmethionine | New England BioLabs | Cat# B9003S |
| [α-32P]-dATP | Perkin elmer | Cat# BLU512H500UC |
| LacI | J. Walter laboratory, (ref. 5) | N/A |
| Bromophenol blue | Pharmacia Biotech | Cat# 17-1329-01 |
| Geminin | J. Walter laboratory, (ref. 41) | N/A |
| Proteinase K | Roche | Cat# 3115879001 |
| RNase A | Sigma | Cat# R4642-250MG |
| Xenopus RTEL1 | J. Walter laboratory, (ref. 14) | N/A |
| MG262 | biotechne | Cat# I-120 |
| 1x cutsmart buffer | New England BioLabs | Cat# B7204 |
| Nt.BspQI | New England BioLabs | Cat# R0644S |
| AatII | New England BioLabs | Cat# R0117L |
| FspI | New England BioLabs | Cat# R0135L |
| Formamide | Roche | Cat# 11814320001 |
| Xylene cyanol FF | Sigma | Cat# X4126 |
| EDTA | Fisher BioReagents | Cat# BP118-500 |
| IgG from non-immunized rabbit serum | Sigma | Cat# R9133-10ML |
| Protein A Sepharose Fast Flow | GE Healthcare | Cat# 17-1279-03 |
| Streptavidin-coupled magnetic beads | Invitrogen | Cat# 11206D |
| Usp21 | D. Finley lab, and (ref. 42) | N/A |
| ANTI-FLAG M2 Affinity Gel | Sigma | Cat# A2220-10ML |
| Nuvia S resin | BioRad | Cat# 1560311 |
| Critical commercial assays | ||
| Q5 site-directed mutagenesis kit | New England BioLabs | Cat# E0554S |
| Capacity cDNA Reverse Transcription Kit | Thermo Fisher | Cat# 4368814 |
| proFIRE Amine Coupling Kit | Dynamic Biosensors | Cat# PF-NH2-1 |
| T-REx Flp-In system | Thermo Fisher | Cat# K650001 |
| alamarBlue assay | Santa Cruz | Cat# sc-206037A |
| NucleoSpin® Gel and PCR Clean-up | MACHERY-NAGEL | Cat# 740609.250 |
| Deposited data | ||
| Original western blot and gel images | This paper; Mendeley Data | doi: 10.17632/spdn6vb4wg.1 |
| Experimental models: Cell lines | ||
| Sf21 cells | Thermo Fisher | Cat# 11497013 |
| U2OS T-REx Flp-In | The Francis Crick Institute Cell Services | N/A |
| Sf9 cells | Expression Systems | Cat# 94-001S |
| Experimental models: Organisms/strains | ||
| Xenopus laevis | Nasco | Cat# LM0053MX |
| Oligonucleotides | ||
| Oligonucleotide sequences used in this study are provided in Table S1 | N/A | N/A |
| Recombinant DNA | ||
| pFastBac1-FANCJ -STREP-WT | This paper | N/A |
| pFastBac1-FANCJ-ZB-STREP-WT | This paper | N/A |
| pFastBac1-FANCJ-ZB-STREP-A349P | This paper | N/A |
| pNIC-HIS-SRAP-WT | This paper | N/A |
| pNIC-HIS-SRAP-C2S | This paper | N/A |
| pNIC-HIS-SRAP-R98E | This paper | N/A |
| p11d-tRPA | Addgene | Cat# 102613 |
| pNIC-STREP-ZB-SPRTN-WT | (ref. 16) | N/A |
| pNIC-STREP-ZB-SPRTN-E112Q | (ref. 16) | N/A |
| pNIC-STREP-ZB-POLη | This paper | N/A |
| pX330-Puro | Addgene | Cat# 82580 |
| pDONR221 | Thermo Fisher | 12536017 |
| pOG44 | Thermo Fisher | Cat# K650001 |
| pcDNA5-FRT/TO-FANCJ-Venus-3xFlag | Addgene | Cat# 17642 |
| pcDNA5-FRT/TO-FANCJ-K52R-Venus-3xFlag | Addgene | Cat# 17643 |
| pcDNA5-FRT/TO-FANCJ-A349P-Venus-3xFlag | This paper | N/A |
| pcDNA5-FRT/TO-Venus-3xFlag-Gateway | Addgene | Cat# 40999 |
| pMBP-HIS-TEV-protease | This paper | N/A |
| pJLS2 | (ref. 14) | N/A |
| pJLS3 | (ref. 14) | N/A |
| pJLS102 | This paper | N/A |
| Software and algorithms | ||
| Prism 7 | GraphPad Software | https://www.graphpad.com/ |
| ImageJ | NIH | https://imagej.net/Fiji/Downloads |
| Affinity Designer | Serif | https://affinity.serif.com/de/designer/#buy |
| Multi-Gauge V3.0 | Fujifilm | N/A |
| Other | ||
| Strep-Tactin® XT 4Flow® | IBA Lifesciences | Cat# 2-5028-001 |
| Superdex® 200 Increase 10/300 GL | GE Healthcare | Cat# GE28-9909-44 |
| HiTrap® Heparin HP | GE Healthcare | Cat# GE17040701 |
| Strep-Tactin® XT Superflow® high-capacity | IBA Lifesciences | Cat# 2-1240-001 |
| HiLoad® 16/600 Superdex® 200 pg | GE Healthcare | Cat# GE28-9893-35 |
| 10 kDa cutoff Amicon Ultra centrifugal filters | Merck | Cat# UFC801096 |
| EconoFit Affi-Gel Blue | BioRad | Cat# 12009234 |
| PD-10 Desalting columns | GE Healthcare | Cat# GE17085101 |