Identifier,Sequence,Purpose,Relevant plasmids
prAH493,gcgctggactgatcaacggc CGAGCTGATCCAAAAAAGAAGAG,Produce PCR fragment for Gibson assembly of 3xNLS[SV40] into PB PGKp-PuroR EF1a 3xFLAG-HaloTag-MCS,PB PGKp-PuroR EF1a 3xFLAG-HaloTag-GDGAGLIN-3xNLS
prAH494,TTGTCGACccaGCGGCCGCG TTCAGGTTCAGGGGGAGG,Produce PCR fragment for Gibson assembly of 3xNLS[SV40] into PB PGKp-PuroR EF1a 3xFLAG-HaloTag-MCS,PB PGKp-PuroR EF1a 3xFLAG-HaloTag-GDGAGLIN-3xNLS
prAH495,AGCT GTGACCGGCGCCTACTCTAG GCCACCATG GCCAGCAACAGCAGCTCC,Produce PCR fragment for Gibson assembly of the RARA coding sequence into XbaI and NotI-H-digested PB PGKp-PuroR EF1a MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR EF1a RARA-HaloTag-GDGAGLIN-3xFLAG
prAH496,ccagcgccatctccgcttGC CGGGGAGTGGGTGGCCG,Produce PCR fragment for Gibson assembly of the RARA coding sequence into XbaI and NotI-H-digested PB PGKp-PuroR EF1a MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR EF1a RARA-HaloTag-GDGAGLIN-3xFLAG
prAH675,GCTTCTGAGGCGGAACGCGAT ATGTACGGGCCAGATATACG,Produce PCR fragment for Gibson assembly of the L30 promoter into AsiSI and XbaI-digested PB PGKp-PuroR EF1a MCS-GDGAGLIN-HaloTag-3xFLAG to produce PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG
prAH676,CGAGccGGTACCGCTAGCTCTAGA GAGGAATTCGCTAGCTCTAG,Produce PCR fragment for Gibson assembly of the L30 promoter into AsiSI and XbaI-digested PB PGKp-PuroR EF1a MCS-GDGAGLIN-HaloTag-3xFLAG to produce PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG
prAH596,AAGCTTGGTACCGAGCTCGGATC GCCACCATGGCCAGCAACAG,Produce PCR fragments for Gibson assembly of RARA[∆DBD] and RARA[∆LBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆DBD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[∆LBD]-GDGAGLIN-HaloTag-3xFLAG
prAH597,gatcagtccagcgccatctccgcttGC,Produce PCR fragments for Gibson assembly of RARA[∆DBD] and RARA[∆LBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆DBD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[∆LBD]-GDGAGLIN-HaloTag-3xFLAG
prAH704,CCTTGGACATGCCCACTTCAAA AGGCTTGTAGATGCGGGGTAG,Produce PCR fragments for Gibson assembly of RARA[∆DBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG to produce PB PGKp-PuroR L30p RARA[∆DBD]-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆DBD]-GDGAGLIN-HaloTag-3xFLAG
prAH705,CTACCCCGCATCTACAAGCCT TTTGAAGTGGGCATGTCCAAGG,Produce PCR fragments for Gibson assembly of RARA[∆DBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆DBD]-GDGAGLIN-HaloTag-3xFLAG
prAH706,GTCCAGGCCCTCTGAGTTCTC CTCCGGCGTCAGCGTGTAG,Produce PCR fragments for Gibson assembly of RARA[∆LBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆LBD]-GDGAGLIN-HaloTag-3xFLAG
prAH707,GCTACACGCTGACGCCGGAG GAGAACTCAGAGGGCCTGGAC,Produce PCR fragments for Gibson assembly of RARA[∆LBD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆LBD]-GDGAGLIN-HaloTag-3xFLAG
prAH1111,GCTAGCGAATTCCTCTCTAG GCCACCATGGCC TGCTTTGTCTGTCAGGACAAGTC,Produce PCR fragment for Gibson assembly of RARA[∆NTD] and RARA[∆NTD ∆CTD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆NTD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[∆NTD ∆CTD]-GDGAGLIN-HaloTag-3xFLAG
prAH1112,catctccgcttGCGGCCGCGGATC C CGGGGAGTGGGTGGCCGG,Produce PCR fragment for Gibson assembly of RARA[∆NTD] and RARA[C88G] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆NTD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[C88G]-GDGAGLIN-HaloTag-3xFLAG
prAH1113,GCTAGCGAATTCCTCTCTAG GCCACCATGGCCAGCAACAGC,Produce PCR fragment for Gibson assembly of RARA[C88G] and RARA[∆CTD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆CTD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[C88G]-GDGAGLIN-HaloTag-3xFLAG
prAH1114,catctccgcttGCGGCCGCGGATC C TGAGTTCTCCAACATTTCCTGG,Produce PCR fragment for Gibson assembly of RARA[∆CTD] and RARA[∆NTD ∆CTD] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[∆CTD]-GDGAGLIN-HaloTag-3xFLAG; PB PGKp-PuroR L30p RARA[∆NTD ∆CTD]-GDGAGLIN-HaloTag-3xFLAG
prAH1069,CTGACAGACAAAGCCAGGCTTGTAGATGCGGGGTAG,Produce PCR fragment for Gibson assembly of RARA[C88G] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[C88G]-GDGAGLIN-HaloTag-3xFLAG
prAH1070,CCCGCATCTACAAGCCTGGCTTTGTCTGTCAGGACAAG,Produce PCR fragment for Gibson assembly of RARA[C88G] into XbaI and BamHI-HF-digested PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG,PB PGKp-PuroR L30p RARA[C88G]-GDGAGLIN-HaloTag-3xFLAG
prAH331,TGT TGG TGA GTG AGC TCT G,Sanger sequencing for vectors descended from PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG; anneals to L30 promoter,
prAH439,ACCGTGCAGGAACAGCAC,Sanger sequencing for vectors descended from PB PGKp-PuroR L30p MCS-GDGAGLIN-HaloTag-3xFLAG; anneals to 5’ end of HaloTag CDS,
prAH1100,CTCTGGGACAAGTTCAGTGAACTC,Sanger sequencing for RARA point mutations and domain deletions; anneals within the RARA ligand-binding domain CDS,
prAH599,CGTTGTAAAACGACGGCCAGT ACACCTGTCCCCATCTGTGTC,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; left homology arm forward,pUC57_homRep_RARA-HaloTag
prAH600,atcagtccagcgccatctcc CGGGGAGTGGGTGGCCG,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; left homology arm reverse,pUC57_homRep_RARA-HaloTag
prAH601,GCCCGGCCACCCACTCCCCG ggagatggcgctggactgat,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; GDGAGLIN-HaloTag-3xFLAG fragment forward,pUC57_homRep_RARA-HaloTag
prAH602,CTGTGTCCATGTGGCGTGGG TTACTTGTCATCGTCATCCTTGTAATC,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; GDGAGLIN-HaloTag-3xFLAG fragment reverse,pUC57_homRep_RARA-HaloTag
prAH603,CAAGGATGACGATGACAAGTAA CCCACGCCACATGGACACA,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; right homology arm forward,pUC57_homRep_RARA-HaloTag
prAH604,CTATGACCATGATTACGCCA CAGGCAGAGGGAGAAAGGG,Produce PCR fragments for Gibson assembly of pUC57_homRep_RARA-HaloTag; right homology arm reverse,pUC57_homRep_RARA-HaloTag
prAH581,CACCG CATGTGGCGTGGGCGGTCAC,Generate pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_1 by Golden Gate cloning,pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_1
prAH609,AAAC GTGACCGCCCACGCCACATG C,Generate pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_1 by Golden Gate cloning,pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_1
prAH583,CACCG CCGTGACCGCCCACGCCACA,Generate pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_2 by Golden Gate cloning,pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_2
prAH610,AAAC TGTGGCGTGGGCGGTCACGG C,Generate pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_2 by Golden Gate cloning,pU6_sgRNA_CBh_Cas9_PGK_Venus_anti-RARA-C_terminus_2
prAH2000,AGTAATAATACGACTCACTATAG,Generate T7-driven templates for sgRNA generation,
prAH2001,AAA AAA GCA CCG ACT CGG TGC CAC TTT TTC AAG TTG ATA ACG GAC TAG CCT TAT TTT AAC TTG CTA TTT CTA GCT CTA AAA C,Generate T7-driven templates for sgRNA generation,ultramer
prAH867,TAGATGTTGAACTATGCAAAGAGACATTTAATTTATTGATGTCTATGAAGTGTTGTGGTTCCTTAACCACATTTCtttttttttttttCCAGGCTATTCAA GAC TTA TGG CAA TGG AGA AAG TCT CTT ggagatggcgctggactgatc,Generate dsDNA homology repair donor for GDGAGLIN-HaloTag-3xFLAG knock-in at human NPM1 C-terminus; ultramer forward ,ultramer
prAH868,GACAACATTTATCAAACACGGTAGGGAAAGTTCTCACTCTGCATTATAAAAAGGACAGCCAGATATCAACTGTTACAGAAATGAAATAAGACGGAAAATTTTTTAACAAATTGTTTAAACTATTTTCTTA CTTGTCATCGTCATCCTTGTAATCG,Generate dsDNA homology repair donor for GDGAGLIN-HaloTag-3xFLAG knock-in at human NPM1 C-terminus; ultramer reverse,ultramer
prAH869,TAGATGTTGAACTATGCAAAGAGACAT,Generate dsDNA homology repair donor for GDGAGLIN-HaloTag-3xFLAG knock-in at human NPM1 C-terminus; outer PCR forward ,
prAH870,GACAACATTTATCAAACACGGTAGG,Generate dsDNA homology repair donor for GDGAGLIN-HaloTag-3xFLAG knock-in at human NPM1 C-terminus; outer PCR reverse,
prAH2000,AGTAATAATACGACTCACTATAG,Generate T7-driven templates for sgRNA generation,
prAH2001,AAA AAA GCA CCG ACT CGG TGC CAC TTT TTC AAG TTG ATA ACG GAC TAG CCT TAT TTT AAC TTG CTA TTT CTA GCT CTA AAA C,Generate T7-driven templates for sgRNA generation,
prAH979,TAATACGACTCACTATAG tCCAGGCTATTCAAGATCTC GTTTTAGAGCTAGAAATAGC,Generate T7-driven template for sgRNA against human NPM1 3’ CDS for GDGAGLIN-HaloTag knock-in; corresponds to sgAH017 (see Guides.csv),
prAH980,TAATACGACTCACTATAG GCCAGAGATCTTGAATAGCC GTTTTAGAGCTAGAAATAGC,Generate T7-driven template for sgRNA against human NPM1 3’ CDS for GDGAGLIN-HaloTag knock-in; corresponds to sgAH018 (see Guides.csv),
prAH761,CGCTAAAGGTCTACGTGCGGA,Genotyping RARA-GDGAGLIN-HaloTag-3xFLAG knock-in; PCR1 (980 bp on WT allele; 1982 bp on correct edited allele),
prAH586,GGGGTCACATGGTCGGTAGAAA,Genotyping RARA-GDGAGLIN-HaloTag-3xFLAG knock-in; PCR1 (980 bp on WT allele; 1982 bp on correct edited allele),
prAH762,ATGGGGGTCGAATGGAAAGCC,Genotyping RARA-GDGAGLIN-HaloTag-3xFLAG knock-in; PCR2 with prAH761 (970 bp on correct edited allele; no expected amplification on WT allele),
prAH763,GCAAGAAGACAACCCGGACCTGAT,Genotyping RARA-GDGAGLIN-HaloTag-3xFLAG knock-in; PCR3 (996 bp on correct edited allele; no expected amplification on WT allele),
prAH764,GTCCTTTCAGGGCTGCCCCACA,Genotyping RARA-GDGAGLIN-HaloTag-3xFLAG knock-in; PCR3 (996 bp on correct edited allele; no expected amplification on WT allele),
prAH1092,CCAACTCTAAAGCTGATCTCGGG,Genotyping NPM1-GDGAGLIN-HaloTag-3xFLAG knock-in cell line; PCR1 and PCR3,
prAH1093,GGAGGGCAGGGACATTCTCATAG,Genotyping NPM1-GDGAGLIN-HaloTag-3xFLAG knock-in cell line; PCR1 and PCR2,
prAH373,CCAAAAGCCTGCCTAACTGC,Genotyping NPM1-GDGAGLIN-HaloTag-3xFLAG knock-in cell line; PCR2,
prAH377,GGGGTCGAATGGAAAGCCA,Genotyping NPM1-GDGAGLIN-HaloTag-3xFLAG knock-in cell line; PCR3,