pMC-TNFalpha
(Plasmid #20210)

• Depositing Lab: Martin Holcik
• Publication: Zhao et al Cell Death Differ. 2009 Feb . 16(2):244-52.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Modifications to backbone: The expression plasmid pMC was constructed by inserting a synthetic polylinker (5′-AGC TTG CGG CCG CTA GCC TCG AGG GAT CCT CTA GAA TGC A-3′) into a pcDNA3 vector (Invitrogen) followed by insertion of the chloramphenicol acetyl transferase (CAT) coding sequence. The CAT insert was generated by PCR amplification using pCAT as a template.
• 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): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TNFa 3' UTR
• Species: H. sapiens (human)
• Insert Size (bp): 808
• Entrez Gene: TNF (a.k.a. DIF, TNF-alpha, TNFA, TNFSF2, TNLG1F)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CAT-F (5'-GTTTGTGATGGCTTCCATGTC-3')

Terms and Licenses

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

Depositor Comments

The 3′UTR was generated by PCR amplification from a liver cDNA library (Invitrogen). and the expression plasmids pMC-TNFα was constructed by inserting the 3′UTR of TNFα downstream of CAT in the pMC construct.