| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| Polyclonal rabbit GFP | Invitrogen/Thermo Fisher | Cat# A11122, RRID:AB_221569 |
| Mouse monoclonal anti-Dlg | Developmental Studies Hybridoma Bank | Cat# 4F3, RRID:AB_528203 |
| Rat monoclonal mCherry | Invitrogen/Thermo Fisher | Cat# M11217, RRID:AB_2536611 |
| Mouse monoclonal dCsp | Developmental Studies Hybridoma Bank | Cat# Ab49, RRID:AB_2307340 |
| Chicken anti-rabbit AlexaFluor 488 | Invitrogen/Thermo-Fisher | Cat# A-21441, RRID:AB_2535859 |
| Goat anti-mouse AlexaFluor 568 | Invitrogen/Thermo-Fisher | Cat# A11004, RRID:AB_2534072 |
| Goat anti-rat AlexaFluor 568 | Invitrogen/Thermo-Fisher | Cat# A-11077, RRID:AB_141874 |
| Chicken anti-mouse AlexaFluor 568 | Invitrogen/Thermo-Fisher | Cat# A21200, RRID:AB_2535786 |
|
| ||
| Bacterial and virus strains | ||
|
| ||
| Chemocompetent Escherichia coli strain DH10B | Invitrogen/Thermo Fisher Scientific | Cat# EC0113 |
| Transformax EPI300 electrocompetent strain cells | Epicenter/Lucigen | Cat# EC300110, |
| EL350 recombineering strain bacterial cells | Kind gift from Donald Court, National Cancer Institute | n/a |
| EC100D pir-116 | Epicenter/Lucigen | Cat# EC6P095H |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| G418 disulfate | VWR | Cat# 97063–060 |
| Puromycin dihydrochloride | VWR | Cat# 97064–280 |
| Blasticidin S hydrochloride | VWR | Cat# 71002–676 |
| Hygromycin B | VWR | Cat# AAJ6068103 |
| Ganciclovir | TCI America | Cat# G0315 |
| 5-Fluorocytosine | TCI America | Cat# F0321 |
| BsaI-HFv2 | New England Biolabs | Cat# R3733 |
| Esp3I | New England Biolabs | Cat# R0734 |
| T4 DNA Ligase | Promega | Cat# M1801 |
| iProof High-Fidelity DNA Polymerase | Biorad | Cat# 1725301 |
| Phusion High-Fidelity DNA Polymerase | New England Biolabs | Cat# M0530 |
| Q5 High-Fidelity DNA Polymerase | New England Biolabs | Cat# M0491 |
| BioReady rTaq DNA polymerase | Bulldog Bio | Cat# BSAX050 |
| Zeocin | Alfa Aesar | Cat# J67140 |
| Phleomycin | VWR | Cat# AAJ67027–8EQ |
| BbsI-HF | New England Biolabs | Cat# R3539L |
| CopyControl Fosmid Autoinduction Solution | Epicenter/Lucigen | Cat# CCIS125 |
| Trypsin protease | Gendepot | Cat# T9600 |
| Ammonium bicarbonate | Fisher Scientific | Cat# 3003–1 |
| LC-MS methanol | Fisher Scientific | Cat# A456 |
| LC-MS water | J.T.Baker | Cat# JT9831–3 |
| LC-MS acetonitrile | Fisher Scientific | Cat# A955 |
| Formic Acid | Fisher Scientific | Cat# A117 |
|
| ||
| Critical commercial assays | ||
|
| ||
| QIAquick PCR Purification Kit | QIAGEN | Cat# 28106 |
| QIAquick Gel Extraction Kit | QIAGEN | Cat# 28506 |
| QIAprep Spin Miniprep Kit | QIAGEN | Cat# 27106 |
| ChargeSwitch-Pro plasmid Miniprep kit | Invitrogen/Thermo Fisher Scientific | Cat# CS30250 |
| ZR-96 Quick-gDNA kit | Zymo Research | Cat# D3010 |
|
| ||
| Experimental models: Organisms/strains | ||
|
| ||
| Drosophila melanogaster: EGFP: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-EGFP-TKpA} VK00033 | This work, available from the BDSC | Cat# 92331, RRID:BDSC_92331 |
| Drosophila melanogaster: G418R: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-nptII-TKpA} VK00033 | This work, available from the BDSC | Cat# 92332, RRID:BDSC_92332 |
| Drosophila melanogaster: PuroR:y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-pac-TKpA} VK00033 | This work, available from the BDSC | Cat# 92333, RRID:BDSC_92333 |
| Drosophila melanogaster: BlastR: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-bsd-TKpA} VK00033/TM6B, Tb[1] | This work, available from the BDSC | Cat# 92334, RRID:BDSC_92334 |
| Drosophila melanogaster: HygroR: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-hph-TKpA} VK00033 | This work, available from the BDSC | Cat# 92335, RRID:BDSC_92335 |
| Drosophila melanogaster: ZeoR: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-bsr-TKpA} VK00033 | This work, available from the BDSC | Cat# 92336, RRID:BDSC_92336 |
| Drosophila melanogaster: GCVS:y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-sr39TK-TKpA} VK00033 | This work, available from the BDSC | Cat# 92337, RRID:BDSC_92337 |
| Drosophila melanogaster: 5FCS: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-Hsp70-CP6-FCU1-TKpA} VK00033 | This work, available from the BDSC | Cat# 92338, RRID:BDSC_92338 |
| Drosophila melanogaster: FM7h-25C-1B: FM7h-RMCE-w[+mC]-25C-1B | This work, available from the BDSC | Cat# 92339, RRID:BDSC_92339 |
| Drosophila melanogaster: CyO-25C-4A: y[1]w[1118]/Dp(1;y)y[+];CyO-RMCE-25C-4A(w[+mC])/L | This work, available from the BDSC | Cat# 92340, RRID:BDSC_92340 |
| Drosophila melanogaster: CyO-52D-3A: y[1]w[1118]/Dp(1;y)y[+];CyO-RMCE-52D-3A(w[+mC])/L | This work, available from the BDSC | Cat# 92341, RRID:BDSC_92341 |
| Drosophila melanogaster: TM6B-25C-5A: y[1]w[1118]/Dp(1;y)y[+];TM6b-RMCE-25C-5A(w[+mC])/D | This work, available from the BDSC | Cat# 92342, RRID:BDSC_92342 |
| Drosophila melanogaster: TM6B-52D-1A: y[1]w[1118]/Dp(1;y)y[+];TM6b-RMCE-52D-1A(w[+mC])/D | This work, available from the BDSC | Cat# 92343, RRID:BDSC_92343 |
| Drosophila melanogaster: TM6bTb::BlastR5FCS: y[1]w[1118]/Dp(1;y)y[+];TM6b-RMCE{BlastR-5FCS-w[5xGMR-CDS]}-25C-5A(w[+mC])/D | This work, available from the BDSC | Cat# 92344, RRID:BDSC_92344 |
| Drosophila melanogaster: TM6bTb::G418R5FCS: y[1]w[1118]/Dp(1;y)y[+];TM6b-RMCE{G418R-5FCS-w[5xGMR-CDS]}-25C-5A(w[+mC])/D | This work, available from the BDSC | Cat# 92345, RRID:BDSC_92345 |
| Drosophila melanogaster: 2xattP::VK00033;VK00020:y[1] M{RFP[3xP3.PB] GFP[E.3xP3] = vas-int.B}ZH-2A w[*]; PBac{y[+]-attP-3B}VK00033, PBac{y[+]-attP-9A}VK00020 | This work, available from the BDSC | Cat# 92346, RRID:BDSC_92346 |
| Drosophila melanogaster: N-EGFP-Csp: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-CH322-06D09-N-EGFP-Csp-G418}VK00033 | This work, available from the BDSC | Cat# 92347, RRID:BDSC_92347 |
| Drosophila melanogaster: NrxIV-CmCherry: y[1]w[1118]; PBac{y[+mDint2] w[+mC] = P[acman]-attB-CH322-154P15-NrxIV-C-Cherry-Blast}VK00033 | This work, available from the BDSC | Cat# 92348, RRID:BDSC_92348 |
| Drosophila melanogaster: R20A02::GAL4; w[5xGMR-CDS] = pR20A02-GAL4-G418R } VK000XX, w[5xGMR-CDS] = p5xUASsfGFP-BlastR }VK000XX 5xUAS::sfGFP: | This work, available from the BDSC | Cat# 92764, RRID:BDSC_92364 |
| Drosophila melanogaster: R76H03::GAL4; 5xUAS::mCherry: w[5xGMR-CDS] = pR76H03-GAL4-G418R }VK000XX, w[5xGMR-CDS] = p5xUAS-mCherry-BlastR } VK000XX | This work, available from the BDSC | Cat# 92765, RRID:BDSC_92365 |
| Drosophila melanogaster: R70B04::LexA; 12xLexAOp::sfGFP: w[+mC] = pR70B04-LexA-G418R }VK000XX, w[+mC] = p12xLexAOp-sfGFP-BlastR }VK000XX | This work, available from the BDSC | Cat# 92766, RRID:BDSC_92366 |
| Drosophila melanogaster: IsoY1: y[1] w[67c23] | This work, available from the BDSC | Cat# 92349, RRID:BDSC_92349 |
|
| ||
| Oligonucleotides | ||
|
| ||
| TKpA-F | This work | N/A |
| TKpA-R | This work | N/A |
| attB-FOR | This work | N/A |
| attB-REV | This work | N/A |
| ModuleA-FOR | This work | N/A |
| ModuleA-REV | This work | N/A |
| ModuleF-FOR | This work | N/A |
| ModuleF-REV | This work | N/A |
| R6Kg-FOR | This work | N/A |
| R6Kg-REV | This work | N/A |
|
| ||
| Recombinant DNA | ||
|
| ||
| p3xP3-Hsp70b | This work, available from AddGene | Cat# 165773 |
| pMCS-TCCC | This work, available from AddGene | Cat# 165774 |
| p5xGMR | This work, available from AddGene | Cat# 165775 |
| pdSCP | This work, available from AddGene | Cat# 165776 |
| pDmActin5c | This work, available from AddGene | Cat# 165777 |
| p3xP3 | This work, available from AddGene | Cat# 165779 |
| pHsp70b-TACT | This work, available from AddGene | Cat# 165780 |
| pDmAlfaTub84b | This work, available from AddGene | Cat# 165781 |
| pDmVasa | This work, available from AddGene | Cat# 165783 |
| pDmHsp70b | This work, available from AddGene | Cat# 165785 |
| pDmPolyUbiq | This work, available from AddGene | Cat# 165786 |
| pPromoterless-P2A | This work, available from AddGene | Cat# 165787 |
| Promoterless TP10 | This work, available from AddGene | Cat# 165790 |
| pR20A02 | This work, available from AddGene | Cat# 165791 |
| pR70B04 | This work, available from AddGene | Cat# 165792 |
| pR76H03 | This work, available from AddGene | Cat# 165793 |
| pCP6-Dros | This work, available from AddGene | Cat# 165794 |
| pCP6-STD | This work, available from AddGene | Cat# 165795 |
| pCP7-STD | This work, available from AddGene | Cat# 165796 |
| pTP10 | This work, available from AddGene | Cat# 165797 |
| pSV40L | This work, available from AddGene | Cat# 165798 |
| pAdh Short 3′UTR | This work, available from AddGene | Cat# 165800 |
| pHSV-TK | This work, available from AddGene | Cat# 165801 |
| prrnBT1_STD | This work, available from AddGene | Cat# 165803 |
| prrnBT2_STD | This work, available from AddGene | Cat# 165804 |
| pL3S2P21_STD | This work, available from AddGene | Cat# 165805 |
| pspy_STD | This work, available from AddGene | Cat# 165806 |
| pGGGSx4 | This work, available from AddGene | Cat# 165807 |
| pT2A | This work, available from AddGene | Cat# 165809 |
| pGGSx4 NT | This work, available from AddGene | Cat# 165810 |
| pGGSx4 CT | This work, available from AddGene | Cat# 165811 |
| p5xUAS-A1 | This work, available from AddGene | Cat# 165812 |
| 4xLexAOp-A1 | This work, available from AddGene | Cat# 165814 |
| 4xLexAOp-A2 | This work, available from AddGene | Cat# 165815 |
| 4xLexAOp-O1 | This work, available from AddGene | Cat# 165816 |
| 4xLexAOp-O2 | This work, available from AddGene | Cat# 165817 |
| 5xQUAS | This work, available from AddGene | Cat# 165818 |
| pGal4BD 1–147 | This work, available from AddGene | Cat# 165819 |
| pGAL4AD 768–881 | This work, available from AddGene | Cat# 165820 |
| pLexA BD | This work, available from AddGene | Cat# 165821 |
| pVP16 AD | This work, available from AddGene | Cat# 165822 |
| pQFBD | This work, available from AddGene | Cat# 165823 |
| pQFAD | This work, available from AddGene | Cat# 165824 |
| pGAL80 | This work, available from AddGene | Cat# 165825 |
| pQS | This work, available from AddGene | Cat# 165826 |
| pmCherry | This work, available from AddGene | Cat# 165828 |
| psfGFP | This work, available from AddGene | Cat# 165829 |
| pEGFP | This work, available from AddGene | Cat# 165830 |
| pEBFP2 | This work, available from AddGene | Cat# 165831 |
| pmCherry TL1 | This work, available from AddGene | Cat# 165832 |
| pmCherry TL5 | This work, available from AddGene | Cat# 165833 |
| pEGFP TL1 | This work, available from AddGene | Cat# 165834 |
| pEGFP TL5 | This work, available from AddGene | Cat# 165835 |
| pNPTII | This work, available from AddGene | Cat# 165836 |
| pPAC | This work, available from AddGene | Cat# 165837 |
| pBSR | This work, available from AddGene | Cat# 165838 |
| pHPH | This work, available from AddGene | Cat# 165839 |
| pBLE | This work, available from AddGene | Cat# 165840 |
| pTK | This work, available from AddGene | Cat# 165841 |
| pFCU1 | This work, available from AddGene | Cat# 165842 |
| pFC31 attB O1 | This work, available from AddGene | Cat# 165843 |
| pFC31 attB O2 | This work, available from AddGene | Cat# 165844 |
| pFC31 | This work, available from AddGene | Cat# 165845 |
| pWhite_CDS | This work, available from AddGene | Cat# 165846 |
| pUPD2 | This work, available from AddGene | Cat# 165856 |
| pVD2_purple | This work, available from AddGene | Cat# 165857 |
| P[acman]-A1 | This work, available from AddGene | Cat# 165858 |
| P[acman]-A2 | This work, available from AddGene | Cat# 165859 |
| P[acman]-O1 | This work, available from AddGene | Cat# 165860 |
| P[acman]-O2 | This work, available from AddGene | Cat# 165861 |
| pR6Kg-A1 | This work, available from AddGene | Cat# 165862 |
| pR6Kg-A2 | This work, available from AddGene | Cat# 165863 |
| pR6Kg-O1 | This work, available from AddGene | Cat# 165864 |
| pR6Kg-O2 | This work, available from AddGene | Cat# 165865 |
| pR6Kg-A1spm | This work, available from AddGene | Cat# 165866 |
| pColE1-AlphaOmega | This work, available from AddGene | Cat# 165867 |
| pEGFP-NT-Tagging | This work, available from AddGene | Cat# 165868 |
| pEGFP-CT-Tagging | This work, available from AddGene | Cat# 165869 |
| pmCherry-NT-Tagging | This work, available from AddGene | Cat# 165870 |
| pmCherry-CT-Tagging | This work, available from AddGene | Cat# 165871 |
| pMiniwhiteA1 | This work, available from AddGene | Cat# 165872 |
| pMiniWhiteA2 | This work, available from AddGene | Cat# 165873 |
| pGMR-WhiteA2 | This work, available from AddGene | Cat# 165874 |
| p3XP3-WhiteA2 | This work, available from AddGene | Cat# 165875 |
| pHSP70B-CP6-nptII | This work, available from AddGene | Cat# 165876 |
| pHSP70B-CP6-pac | This work, available from AddGene | Cat# 165877 |
| pHSP70B-CP6-bsr | This work, available from AddGene | Cat# 165878 |
| pHSP70B-CP6-hph | This work, available from AddGene | Cat# 165879 |
| pHSP70B-CP6-ble | This work, available from AddGene | Cat# 165880 |
| pHSP70B-CP6-TK | This work, available from AddGene | Cat# 165881 |
| pHSP70B-CP6-FCU1 | This work, available from AddGene | Cat# 165882 |
| pG418R-GMRWhite | This work, available from AddGene | Cat# 165883 |
| pPuroR-GMRWhite | This work, available from AddGene | Cat# 165884 |
| pBlastR-GMRWhite | This work, available from AddGene | Cat# 165885 |
| pHygroR-GMRWhite | This work, available from AddGene | Cat# 165886 |
| pRMCE{G418R-5FCS-GMRWhite}-VasaphiC31 | This work, available from AddGene | Cat# 165887 |
| pRMCE{PuroR-5FCS-GMRWhite}-VasaphiC31 | This work, available from AddGene | Cat# 165888 |
| pRMCE{BlastR-5FCS-GMRWhite}-VasaphiC31 | This work, available from AddGene | Cat# 165889 |
| pG418R-MCS-GAL4-MiniWhite | This work, available from AddGene | Cat# 165890 |
| pPuroR-MCS-GAL4-MiniWhite | This work, available from AddGene | Cat# 165892 |
| pBlastR-MCS-GAL4-MiniWhite | This work, available from AddGene | Cat# 165894 |
| pG418R-MCS-LexA-VP16-GMRWhite | This work, available from AddGene | Cat# 165895 |
| pG418R-MCS-LexA-VP16-MiniWhite | This work, available from AddGene | Cat# 165896 |
| pPuroR-MCS-LexA-VP16-GMRWhite | This work, available from AddGene | Cat# 165897 |
| pPuroR-MCS-LexA-VP16-MiniWhite | This work, available from AddGene | Cat# 165898 |
| pBlastR-MCS-LexA-VP16-MiniWhite | This work, available from AddGene | Cat# 165900 |
| pG418R-MCS-QF-MiniWhite | This work, available from AddGene | Cat# 165902 |
| pBlastR-MCS-QF-MiniWhite | This work, available from AddGene | Cat# 165906 |
| pBlastR-5xUAS-sfGFP-GMRWhite | This work, available from AddGene | Cat# 165907 |
| pBlastR-5xUAS-mCherry-GMRWhite | This work, available from AddGene | Cat# 165908 |
| pBlastR-5xUAS-sfGFP-MiniWhite | This work, available from AddGene | Cat# 165910 |
| pBlastR-5xUAS-mCherry-MiniWhite | This work, available from AddGene | Cat# 165911 |
| pBlastR-5xUAS-EBFP2-MiniWhite | This work, available from AddGene | Cat# 165912 |
| pBlastR-12xLexAOp-sfGFP-MiniWhite | This work, available from AddGene | Cat# 165916 |
| pBlastR-12xLexAOp-mCherry-MiniWhite | This work, available from AddGene | Cat# 165917 |
| pBlastR-12xLexAOp-EBFP2-MiniWhite | This work, available from AddGene | Cat# 165918 |
| pBlastR-5xQUAS-mCherry-GMRWhite | This work, available from AddGene | Cat# 165920 |
| pBlastR-5xQUAS-EBFP2-GMRWhite | This work, available from AddGene | Cat# 165921 |
| pBlastR-5xQUAS-sfGFP-MiniWhite | This work, available from AddGene | Cat# 165922 |
| pBlastR-5xQUAS-mCherry-MiniWhite | This work, available from AddGene | Cat# 165923 |
| pBlastR-5xQUAS-EBFP2-MiniWhite | This work, available from AddGene | Cat# 165924 |
|
| ||
| Software and algorithms | ||
|
| ||
| Prism 7 software v9.1.1 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| Adobe Illustrator Creative Cloud | Adobe | https://www.adobe.com/creativecloud.html |
| Adobe Photoshop Creative Cloud | Adobe | https://www.adobe.com/creativecloud.html |
| Search Tool for the Retrieval of Interacting Genes/Proteins | STRING (https://www.string-db.org) | Szklarczyk et al., 2019 |
| Cytoscape v3.8.2 | https://cytoscape.org | Shannon et al., 2003 |
| Zen Software Blue Version 2.3 pro HWL | Zeiss | https://www.zeiss.com/microscopy/us/products/microscope-software/zen.html |
| Codon Optimization Tool | IDT | http://www.idtdna.com/pages/tools/codon-optimization-tool?returnurl=%2FCodonOpt |
| Splice Site Prediction by Neural Network | Berkley Drosophila Genome Project | https://www.fruitfly.org/seq_tools/splice.html |
| SnapGene v4.2.3 | GSL Biotech LLC | https://www.snapgene.com:443/products/snapgene/ |
|
| ||
| Other | ||
|
| ||
| ChromoTek GFP-Trap Dynabeads | ChromoTek | Cat# gtd-10, RRID:AB_2827592 |
| NuPAGE 10%, Bis-Tris protein gel | Thermo Fisher Scientific | NP0315BOX |
| Reprosil-Pur Basic C18 beads | Dr.Maisch GmbH, Germany | R119.b9.3 |
| Fused silica tubing | IDEX Health | FS-110 |
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