pRSET-4xTR-TUBE
(Plasmid #110312)

• Purpose: E. coli expression and purification of 4xTR-TUBE
• Depositing Lab: Yasushi Saeki
• Publication: Tsuchiya et al Nat Commun. 2018 Feb 6;9(1):524. doi: 10.1038/s41467-018-02869-x.

Backbone

• Vector backbone: pRSET-A
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2685
• Total vector size (bp): 3519
• Modifications to backbone: In-Fusion cloning was used for construction of this plasmid.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: TR-TUBE was expressed in E.coli Rosetta2 (DE3) with 0.2 mM IPTG for 15 h at 22°C.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 4xTR-TUBE
• Alt name: 4 tandem repeats of the trypsin-resistant UBQLN1 UBA domain
• Species: H. sapiens (human)
• Insert Size (bp): 834
• Mutation: All Arg residues in the UBA domain are mutated to Ala.
• GenBank ID: NM_013438.4
• Entrez Gene: UBQLN1 (a.k.a. DA41, DSK2, PLIC-1, UBQN, XDRP1)
• Promoter: T7
• 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

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Nucleotide sequence is not identical to NM_013438.4, but amino acid sequence is as described in Gene/Insert: Mutation.