pET28b_Q69L
(Plasmid #186272)

• Purpose: Expression vector of His-RanQ69L in E.coli
• Depositing Lab: Masahito Ikawa
• Publication: Mori et al J Cell Biol. 2021 Oct 4;220(10). pii: 212591. doi: 10.1083/jcb.202012001. Epub 2021 Aug 23.

Backbone

• Vector backbone: pET28b
• Backbone size w/o insert (bp): 5368
• Total vector size (bp): 5978
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: GTP-binding nuclear protein Ran
• Alt name: Ran
• Species: M. musculus (mouse)
• Insert Size (bp): 651
• Mutation: Changed Q69 into L
• GenBank ID: NM_009391
• Entrez Gene: Ran
• Promoter: T7 promoter
• Tag / Fusion Protein:
  • His-tagged at N terminal (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: TAATA CGACT CACTA TAGGG
• 3′ sequencing primer: ATGCT AGTTA TTGCT CAGCG G

Terms and Licenses

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

Depositor Comments

For protein expression, transform into BL21(DE3) E. coli because the pET vector system requires T7 RNA polymerase for expression. Induce with 1 mM ITPG for 2h at 28 °C.