pcDNA3-hsGFP
(Plasmid #64912)

• Purpose: Encodes a Fluorescent Reporter for Hydrogen Sulfide in Mammalian Cells
• Depositing Lab: Huiwang Ai
• Publication: Chen et al Biochemistry. 2014 Sep 23;53(37):5966-74. doi: 10.1021/bi500830d. Epub 2014 Sep 5.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5968
• Total vector size (bp): 6748
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: hsGFP
• Species: Synthetic
• Insert Size (bp): 780
• Mutation: Chromophore tyrosine of the protein mutated to TAG amber codon
• Promoter: CMV
• Tag / Fusion Protein:
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: pCMV-F
• 3′ sequencing primer: SP6

Terms and Licenses

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

Depositor Comments

Please cite Z. Chen and H-w. Ai*, Biochemistry, 2014, 53: 5966-5974; and S. Chen, Z. Chen, W. Ren and H-w. Ai*, J. Am. Chem. Soc., 2012, 134: 9589-9592. Please note that you will also need a pMAH plasmid for mammalian expression. pMAH-POLY (Z. Chen, W. Ren, Q.E. Wright and H-w. Ai*, J. Am. Chem. Soc., 2013, 135: 14940-14943.) is also deposited to Addgene, which can be used to replace pMAH-AzF for mammalian applications.