pMAH-POLY
(Plasmid #64915)

• Purpose: Encodes a polyspecific aminoacyl-tRNA synthetase and four copies of amber suppression tRNAs for unnatural amino acid incorporation in mammalian cells
• Depositing Lab: Huiwang Ai
• Publication: Chen et al J Am Chem Soc. 2013 Oct 9;135(40):14940-3. doi: 10.1021/ja408011q. Epub 2013 Sep 26.

Backbone

• Vector backbone: pMAH
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 8244
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH10B
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Polyspecific aminoacyl-tRNA Synthetase
• Alt name: E. coli tyrosyl synthetase with Y37I/D182S/F183M/D265R mutations
• Species: Synthetic
• Insert Size (bp): 1272
• Promoter: CMV
• Tag / Fusion Protein:
  • None

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: PmeI or ApaI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-R

Gene/Insert 2

• Gene/Insert name: Amber Suppression tRNA
• Species: Synthetic
• Promoter: U6/H1

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: SalI (destroyed during cloning)
• 5′ sequencing primer: tRNA-F
• 3′ sequencing primer: tRNA-R

Terms and Licenses

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

Depositor Comments

Please cite Z. Chen, W. Ren, Q.E. Wright and H-w. Ai*, J. Am. Chem. Soc., 2013, 135: 14940-14943.