pET30a-AlkB-D135S
(Plasmid #79051)

• Purpose: Express mutant AlkB protein (D135S) in E. coli BL21(DE3)
• Depositing Lab: Tao Pan
• Publication: Zheng et al Nat Methods. 2015 Sep;12(9):835-7. doi: 10.1038/nmeth.3478. Epub 2015 Jul 27.

Backbone

• Vector backbone: pET30a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5243
• Total vector size (bp): 5858
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: BL21 (DE3) for expression
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: AlkB
• Species: E.coli
• Insert Size (bp): 615
• Mutation: A truncated AlkB with deletion of the amino (N)-terminal 11 amino acids and D135S mutation
• GenBank ID: 946708
• Entrez Gene: alkB (a.k.a. b2212, ECK2204, aidD)
• Promoter: T7
• Tag / Fusion Protein:
  • His tag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (destroyed during cloning)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7-term

Resource Information

• Articles Citing this Plasmid:
  • 9 References

Terms and Licenses

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