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PurposeE. coli expression and purification of 6xTR-TUBE
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 110313 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRSET-A
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 2685
- Total vector size (bp) 3891
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Modifications to backboneIn-Fusion cloning was used for construction of this plasmid.
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsTR-TUBE was expressed in E.coli Rosetta2 (DE3) with 0.2 mM IPTG for 15 h at 22°C.
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert name6xTR-TUBE
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Alt name6 tandem repeats of the trypsin-resistant UBQLN1 UBA domain
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SpeciesH. sapiens (human)
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Insert Size (bp)1206
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MutationAll Arg residues in the UBA domain are mutated to Ala.
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GenBank IDNM_013438.4
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Entrez GeneUBQLN1 (a.k.a. DA41, DSK2, PLIC-1, UBQN, XDRP1)
- Promoter T7
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Tag
/ Fusion Protein
- N-terminal His6-T7 tag (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer TAATACGACTCACTATAGG
- 3′ sequencing primer CTAGTTATTGCTCAGCGG (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
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
Nucleotide sequence is not identical to NM_013438.4, but amino acid sequence is as described in Gene/Insert: Mutation.
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:
pRSET-6xTR-TUBE was a gift from Yasushi Saeki (Addgene plasmid # 110313 ; http://n2t.net/addgene:110313 ; RRID:Addgene_110313) -
For your References section:
Ub-ProT reveals global length and composition of protein ubiquitylation in cells. Tsuchiya H, Burana D, Ohtake F, Arai N, Kaiho A, Komada M, Tanaka K, Saeki Y. Nat Commun. 2018 Feb 6;9(1):524. doi: 10.1038/s41467-018-02869-x. 10.1038/s41467-018-02869-x [pii] PubMed 29410401