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pCX-TRI-2A
(Plasmid #74674)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 74674 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCX-2A-2A
  • Backbone manufacturer
    Giovannoni lab, Addgene plasmid 74670
  • Total vector size (bp) 10110
  • 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
    HO1
  • Species
    H. sapiens (human)
  • Entrez Gene
    HMOX1 (a.k.a. HMOX1D, HO-1, HSP32, bK286B10)
  • 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 NheI (not destroyed)
  • 5′ sequencing primer pCAX-F CAGCTCCTGGGCAACGTGC
  • (Common Sequencing Primers)

Gene/Insert 2

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

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site AflII (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer none
  • (Common Sequencing Primers)

Gene/Insert 3

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

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site XhoI (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer M13R
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    HMOX1 (hHO-1) sequence was kindly provided by Prof. Fritz Bach's laboratory at Harvard Medical School, Boston. 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
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Contains a D473N mutation in CD73, but it does not impair either the plasmid function or the hCD73 protein activity.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCX-TRI-2A was a gift from Roberto Giovannoni (Addgene plasmid # 74674 ; http://n2t.net/addgene:74674 ; RRID:Addgene_74674)
  • For your References section:

    Simultaneous Overexpression of Functional Human HO-1, E5NT and ENTPD1 Protects Murine Fibroblasts against TNF-alpha-Induced Injury In Vitro. Cinti A, De Giorgi M, Chisci E, Arena C, Galimberti G, Farina L, Bugarin C, Rivolta I, Gaipa G, Smolenski RT, Cerrito MG, Lavitrano M, Giovannoni R. PLoS One. 2015 Oct 29;10(10):e0141933. doi: 10.1371/journal.pone.0141933. eCollection 2015. PONE-D-15-33437 [pii] PubMed 26513260