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PurposeCRBN midi construct for use in crystallography and biophysical studies enabling ligand and degrader design
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 215330 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepNIC28-Bsa4
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 7280
- Total vector size (bp) 8318
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Modifications to backbonenone
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsBL21 (DE3) for bacterial expression of encoding protein
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameCRBN
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SpeciesH. sapiens (human)
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Insert Size (bp)1038
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Mutationcontains amino acids 41-187 followed by a Gly-Ser-Gly linker and amino acids 249-426, with mutations C78I, I92V, K116N, Q134E, R283W, C287N, V293S, G302D, L342R, C343E, T359I and L423I
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GenBank IDNM_016302.4
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Entrez GeneCRBN (a.k.a. MRT2, MRT2A)
- Promoter lacI
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Tags
/ Fusion Proteins
- 6xHis (N terminal on backbone)
- TEV (N terminal on backbone)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer T7
- 3′ sequencing primer T7 terminator (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Please visit https://doi.org/10.1101/2024.01.17.575503 for bioRxiv preprint.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
His6-TEV-CRBN-midi was a gift from Alessio Ciulli (Addgene plasmid # 215330 ; http://n2t.net/addgene:215330 ; RRID:Addgene_215330) -
For your References section:
Design of a Cereblon construct for crystallographic and biophysical studies of protein degraders. Kroupova A, Spiteri VA, Rutter ZJ, Furihata H, Darren D, Ramachandran S, Chakraborti S, Haubrich K, Pethe J, Gonzales D, Wijaya AJ, Rodriguez-Rios M, Sturbaut M, Lynch DM, Farnaby W, Nakasone MA, Zollman D, Ciulli A. Nat Commun. 2024 Oct 15;15(1):8885. doi: 10.1038/s41467-024-52871-9. 10.1038/s41467-024-52871-9 PubMed 39406745