pCX-DI-2A
(Plasmid #74671)

• Purpose: Mammalian expression of DI-2A (hCD73/F2A/hCD39)
• Depositing Lab: Roberto Giovannoni
• Publication: De Giorgi et al Nucleosides Nucleotides Nucleic Acids. 2014;33(4-6):313-8. doi: 10.1080/15257770.2014.896466.

Backbone

• Vector backbone: pCX-2A-2A
• Backbone manufacturer: Giovannoni lab, Addgene plasmid 74670
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: CD73
• Species: H. sapiens (human)
• Entrez Gene: NT5E (a.k.a. CALJA, CD73, E5NT, NT, NT5, NTE, eN, eNT)
• Promoter: pCAGGS
• Tag / Fusion Protein:
  • F2A (C terminal on backbone)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: pCAX-F CAGCTCCTGGGCAACGTGC

Gene/Insert 2

• Gene/Insert name: CD39
• Species: H. sapiens (human)
• Entrez Gene: ENTPD1 (a.k.a. ATPDase, CD39, NTPDase-1, SPG64)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: unknown
• 3′ sequencing primer: M13R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: NT5E (hCD73) sequence was kindly provided by Dr. Jozef Spychala at University of Michigan, Ann Arbor. ENTPD1 (hCD39) sequence was kindly provided by Dr. Simon Robson at Harvard Medical School, Boston.

Terms and Licenses

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

Depositor Comments

Additional details can be found in the following publication:
Simultaneous overexpression of human E5NT and ENTPD1 protects porcine endothelial cells against H2O2-induced oxidative stress and cytotoxicity in vitro. Free Radic Biol Med. 2017 Jul;108:320-333. doi: 10.1016/j.freeradbiomed.2017.03.038.
PMID: 28389406