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pAAV-Ptet-RFP-shR-rtTA
(Plasmid #35625)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 35625 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAAV with pTRIPZ elements
  • Backbone manufacturer
    Stratagene and Open Biosystems
  • Backbone size (bp) 7498
  • Modifications to backbone
    A 1.3-kb filler DNA fragment flanked by two BspQ1 restriction enzyme sites was inserted between the 5′mir and the 3′mir sequences. Once digested with BspQ1, the double-digested vector then has two incompatible 5′ overhang ends and can be easily isolated from any single digested vector, thus eliminating self-ligation background in later procedures. The digested vector is ready for the insertion of the shRNA sequence, which is generated by annealing two single-stranded oligonucleotides to form a small double-stranded insert with proper overhangs as described in the paper.
  • Vector type
    Mammalian Expression, AAV, RNAi ; Tet inducible
  • Promoter Ptet

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    Stbl3
  • Copy number
    Unknown

Depositor Comments

This plasmid is meant to be used in combination with pGFPns-reporter (Addgene #35626) to test the efficacy of shRNAs.

The vector is 8756bp including a 1258bp filler, after cutting with BspQ1 the vector is 7498bp.

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:

    pAAV-Ptet-RFP-shR-rtTA was a gift from Howard Gu (Addgene plasmid # 35625 ; http://n2t.net/addgene:35625 ; RRID:Addgene_35625)
  • For your References section:

    Fluorescence-based evaluation of shRNA efficacy. Naughton BJ, Han DD, Gu HH. Anal Biochem. 2011 Oct 1;417(1):162-4. Epub 2011 Jun 13. 10.1016/j.ab.2011.06.008 PubMed 21726522