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p201N 1514
(Plasmid #47025)

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Item Catalog # Description Quantity Price (USD)
Plasmid 47025 Standard format: Plasmid sent in bacteria as agar stab 1 $85 *
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Backbone

  • Vector backbone
    pPZP
  • Backbone size w/o insert (bp) 6200
  • Total vector size (bp) 10626
  • Modifications to backbone
    Inclusion of aph gene for bacterial selection on kanamycin.
  • Vector type
    RNAi
  • Selectable markers
    G418, kanamycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH10B
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    gma-miR1514 recognition sequence
  • Species
    Glycine max
  • Insert Size (bp)
    22
  • Promoter GmUbi

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site AscI (destroyed during cloning)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer CGAGATTGCTTCAGATCCGTA
  • 3′ sequencing primer CCATTTCCATTTCACAGTTCG
    • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    nptII selectable marker
  • Alt name
    neomycin phosphotransferase II
  • Species
    Escherichia coli
  • Insert Size (bp)
    795
  • Promoter StUbi-3

Cloning Information for Gene/Insert 2

  • Cloning method Unknown
  • 5′ sequencing primer AGCGGATAACAATTTCACACAGGA
  • 3′ sequencing primer GCAGAGCTTACACTCTCATTC
    • (Common Sequencing Primers)

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The GmUbi promoter requires a license for distribution to commercial entities.

miRNA target sequence is: CAATGCCTATTTTAGAAATGAA

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:

    p201N 1514 was a gift from Wayne Parrott (Addgene plasmid # 47025 ; http://n2t.net/addgene:47025 ; RRID:Addgene_47025)

  • For your References section:

    Simple gene silencing using the trans-acting siRNA pathway. Jacobs TB, Lawler NJ, LaFayette PR, Vodkin LO, Parrott WA. Plant Biotechnol J. 2015 Mar 27. doi: 10.1111/pbi.12362. 10.1111/pbi.12362 PubMed 25816689
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