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pCX-DI-2A
(Plasmid #74671)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 74671 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 74671-D50 50 μg of DNA in Tris buffer $495
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DNA 74671-D100 100 μg of DNA in Tris buffer $585
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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
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    CD39
  • Species
    H. sapiens (human)
  • Entrez Gene
    ENTPD1 (a.k.a. ATP-DPH, 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
  • (Common Sequencing Primers)

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
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

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

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

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-DI-2A was a gift from Roberto Giovannoni (Addgene plasmid # 74671 ; http://n2t.net/addgene:74671 ; RRID:Addgene_74671)

  • For your References section:

    Functional analysis of expression of human ecto-nucleoside triphosphate diphosphohydrolase-1 and/or ecto-5'-nucleotidase in pig endothelial cells. De Giorgi M, Pelikant-Malecka I, Sielicka A, Slominska EM, Giovannoni R, Cinti A, Cerrito MG, Lavitrano M, Smolenski RT. Nucleosides Nucleotides Nucleic Acids. 2014;33(4-6):313-8. doi: 10.1080/15257770.2014.896466. 10.1080/15257770.2014.896466 PubMed 24940685
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