Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | KIAA1279 | GenBank | HGNC:23419 | |
| Gene (Mus musculus) | KIF1A | GenBank | MGI:108391 | |
| Gene (Mus musculus) | KIF15 | GenBank | MGI:1098258 | |
| Strain, strain background (Escherichia coli) | BL21(DE3) | NEB | Cat. #: C2527H | Competent cells |
| Strain, strain background (Escherichia coli) | BL21-Gold (DE3) | Agilent | Cat. #: 230130 | Competent cells |
| Strain, strain background (Escherichia coli) | Rosetta2 (DE3) | Novagen | Cat. #: 71400 | Competent cells |
| Cell line (Homo sapiens) | Human embryonic kidney 239T (HEK293T) | ATCC | CRL-3216 RRID:CVCL_0063 | |
| Cell line (Cercopithecus aethiops) | Cercopithecus aethiops kidney (COS-7) | ATCC | CRL-1651 RRID:CVCL_0224 |
|
| Peptide, recombinant protein | Porcine tubulin (>99% pure) | Cytoskeleton Inc | Cat. #: T240 | |
| Antibody | Anti-HA (mouse monoclonal) |
Roche | Cat# 11666606001; RRID:AB_514506 | IF (1:500) |
| Antibody | Anti-mouse IgG1, Alexa488 (goat polyclonal) |
Thermo Fisher Scientific | Cat# A-21121, RRID:AB_2535764 | IF (1:400) |
| Antibody | Anti-HA (mouse monoclonal) | Biolegend | Cat# 901533; RRID:AB_2801249 |
WB (1:2000) |
| Antibody | Anti-GFP (rabbit polyclonal) | Abcam | Cat# ab290; RRID:AB_303395 | WB (1:10000) |
| Antibody | Anti-rabbit IgG antibody, IRDye 680LTconjugated (goat polyclonal) | LI-COR Biosciences | Cat# 827–11081; RRID:AB_10795015 | WB (1:20000) |
| Antibody | Anti-mouse IgG antibody, IRDye 800CWconjugated (goat polyclonal) |
LI-COR Biosciences | Cat# 827–08364; RRID:AB_10793856 | WB (1:15000) |
| Recombinant DNA reagent | KBP (plasmid) | Kevenaar et al., 2016 | Described in Materials and methods | |
| Recombinant DNA reagent | KIF1A_MD (plasmid) | Atherton et al., 2014 | Described in Materials and methods | |
| Recombinant DNA reagent | KIF15_MD (plasmid) | This study | Described in Materials and methods | |
| Recombinant DNA reagent | pebioGFP (plasmid) | van der Vaart et al., 2013 | N/A | Described in Materials and methods |
| Recombinant DNA reagent | BirA coding vector (plasmid) | van der Vaart et al., 2013 | N/A | Described in Materials and methods |
| Recombinant DNA reagent | GW1–PEX3–mRFP–FKBP1 (plasmid) |
Kevenaar et al., 2016 | N/A | Described in Materials and methods |
| Recombinant DNA reagent | β-actin–Kif1A_MDC–FRB (plasmid) |
Kevenaar et al., 2016 | N/A | Described in Materials and methods |
| Recombinant DNA reagent | β-actin–Kif15_MDC–FRB (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pebioGFP-Kif1A_MDC (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pebioGFP-Kif15_MDC (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L1 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L3 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L5 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L10 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L12 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L14 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L16 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L18 (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L10+L12 (plasmid) | This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L10+L14 (plasmid) | This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_L12+L14 (plasmid) | This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_αHP4a (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_αHP4b (plasmid) |
This study | N/A | Described in Materials and methods |
| Recombinant DNA reagent | pGW1–HA–KBP_αHP5a (plasmid) |
This study | N/A | Described in Materials and methods |
| Sequence-based reagent | KBP_fwd | This study | PCR primer for KBP mutants | TATTATTATGGCGCGCCAGGATCCCCGGAATTCGGCACGAGGGAGGCCGCTATGGCGAACGTTCCGTGGGCA |
| Sequence-based reagent | KBP_rev | This study | PCR primer for KBP mutants | CTCGTCGACTCCTAATCCTTAAGTCAGGGCCATCTT |
| Sequence-based reagent | KBP_L1_fwd | This study | PCR primer for KBP_L1 | CTGCATAAAAATCCGGCAGCAGCACCAGCAGCATCCAAATACAGCGCC |
| Sequence-based reagent | KBP_L1_rev | This study | PCR primer for KBP_L1 | GGCGCTGTATTTGGATGCTGCTGGTGCTGCTGCCGGATTTTTATGCAG |
| Sequence-based reagent | KBP_L3_fwd | This study | PCR primer for KBP_L3 | TGAACCACATCGACGCAGGAGGACTGTCGGCGGGGGA |
| Sequence-based reagent | KBP_L3_rev | This study | PCR primer for KBP_L3 | TCCCCCGCCGACAGTCCTCCTGCGTCGATGTGGTTCA |
| Sequence-based reagent | KBP_L5_fwd | This study | PCR primer for KBP_L5 | ATCTTGTGGTCTGAAGCAGGAGCAATTGAAACTGCACAG |
| Sequence-based reagent | KBP_L5_rev | This study | PCR primer for KBP_L5 | CTGTGCAGTTTCAATTGCTCCTGCTTCAGACCACAAGAT |
| Sequence-based reagent | KBP_L10_fwd | This study | PCR primer for KBP_L10 | TTTGGTCAAACTGGAGCAGGAGCAGGAGCAGGAGCAGGACCAGCAGGAGCAGGAGCAGGACCAGGAGGATATCATCAAAGAAA |
| Sequence-based reagent | KBP_L10_rev | This study | PCR primer for KBP_L10 | TTTCTTTGATGATATCCTCCTGGTCCTGCTCCTGCTCCTGCTGGTCCTGCTCCTGCTCCTGCTCCTGCTCCAGTTTGACCAAA |
| Sequence-based reagent | KBP_L12_fwd | This study | PCR primer for KBP_L12 | GAGTTCTTTCAGATTGGCGGCGCGGTCACTGACCATATT |
| Sequence-based reagent | KBP_L12_rev | This study | PCR primer for KBP_L12 | AATATGGTCAGTGACCGCGCCGCCAATCTGAAAGAACTC |
| Sequence-based reagent | KBP_L14_fwd | This study | PCR primer for KBP_L14 | TAGAGCCCCTAACTGTAGCAGCAGGACCAGCAGCATATCTGTTGGTCAAC |
| Sequence-based reagent | KBP_L14_rev | This study | PCR primer for KBP_L14 | GTTGACCAACAGATATGCTGCTGGTCCTGCTGCTACAGTTAGGGGCTCTA |
| Sequence-based reagent | KBP_L16_fwd | This study | PCR primer for KBP_L16 | TCCCTGAGAGACCCAGCAGCAGGAGCACCAGCAGGAGCAGGAGCAGGAGCAGCACGCCCTGCCATGTTA |
| Sequence-based reagent | KBP_L16_rev | This study | PCR primer for KBP_L16 | TAACATGGCAGGGCGTGCTGCTCCTGCTCCTGCTCCTGCTGGTGCTCCTGCTGCTGGGTCTCTCAGGGA |
| Sequence-based reagent | KBP_L18_fwd | This study | PCR primer for KBP_L18 |
ATTGTTGATTACTGTGCAGCAGGACCAGGAGCCGCCCAGGAAATA |
| Sequence-based reagent | KBP_L18_rev | This study | PCR primer for KBP_L18 | TATTTCCTGGGCGGCTCCTGGTCCTGCTGCACAGTAATCAACAAT |
| Sequence-based reagent | KBP_HP4a_fwd | This study | PCR primer for KBP_L HP4a | ACTCATAACCTATATGCACTAGCTGCAGTCTACCAGCATCTG |
| Sequence-based reagent | KBP_L HP4a _rev | This study | PCR primer for KBP_ HP4a | CAGATGCTGGTAGACTGCAGCTAGTGCATATAGGTTATGAGT |
| Sequence-based reagent | KBP_HP4b_fwd | This study | PCR primer for KBP_L HP4b | AGTACACTAAAACGCGCACTTGAGCACAATGCC |
| Sequence-based reagent | KBP_L HP4b _rev | This study | PCR primer for KBP_ HP4b | GGCATTGTGCTCAAGTGCGCGTTTTAGTGTACT |
| Sequence-based reagent | KBP_HP5a_fwd | This study | PCR primer for KBP_L HP5a | GCTATCAATGCTGCTGCGTTGTCAGCGTTTTACATCAATAAG |
| Sequence-based reagent | KBP_ HP5a _rev | This study | PCR primer for KBP_ HP5a | CTTATTGATGTAAAACGCTGACAACGCAGCAGCATTGATAGC |
| Sequence-based reagent | KIF15_FRB_fwd | This study | PCR primer for KIF15–FRB | AAGCTTGCCACCATGGGCGCGCCTGCCACCATGGCTCCTGGCTGCAAATCT |
| Sequence-based reagent | KIF15_FRB_rev | This study | PCR primer for KIF15–FRB | AGAGGATTCTAGAAGCAGGCGCGCCAGCGTAGTCTGGGACGTCGTATGGGTAGAATTCTCCTGGTGTCAGCTGCCCAGA |
| Sequence-based reagent | bioGFPKIF15 _fwd | This study | PCR primer for bioGFPKIF15 | AGCTCAAGCTTCGAATTGGGCGCGCCAGCCACCATGGCTCCTGGCTGCAAATCT |
| Sequence-based reagent | bioGFPKIF15_rev | This study | PCR primer for bioGFPKIF15 | GAATTCGATATCCTGCAGGTCGACTCCAGATCCTCATCCTGGTGTCAGCTGCCCAGA |
| Sequence-based reagent | bioGFPKIF1A _fwd | This study | PCR primer for bioGFPKIF1A | TATTATAATGGCGCGCCAGCCACCGCCGGGGCCTCTGTGAAGGT |
| Sequence-based reagent | bioGFPKIF1A_rev | This study | PCR primer for bioGFPKIF1A | CTCGTCGACTCCTCCTCCTCATTTGGGAGAAAACACACCCAA |
| Commercial assay or kit | EnzChek Phosphate Assay Kit | Invitrogen | E6646 | |
| Chemical compound, drug | AP21967 | TaKaRa | Cat# 635057 | 1 µM |
| Chemical compound, drug | PEI | PolySciences | Cat# 24765–2 | |
| Chemical compound, drug | Fugene | Promega | Cat# E2692 | |
| Software, algorithm | ImageJ | NIH | https://imagej.nih.gov/ij/; RRID:SCR_003070 | |
| Software, algorithm | RELION | Zivanov et al., 2018 | n/a | |
| Software, algorithm | CryoSparc2 | Punjani et al., 2017 | n/a | |
| Software, algorithm | CisTEM | Grant et al., 2018 | n/a | |
| Software, algorithm | MiRP | Cook et al., 2020 | n/a | Protocol implemented in RELION |