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

• Purpose: (Empty Backbone)
• Depositing Lab: Howard Gu
• Publication: Naughton et al Anal Biochem. 2011 Oct 1;417(1):162-4. Epub 2011 Jun 13.

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, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Resource Information

• Supplemental Documents:
  • pAAV-Ptet-RFP-shR-rtTA.pdf
• Addgene Notes:
  • Addgene Diagnostic Digest- EcoRI, BspQI
  • Addgene Diagnostic Digest- HpaI, HpaI/AgeI
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

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.

Note: Addgene NGS results identified an insertion in the backbone, that makes the total size 9112. Plasmid is still predicted to function as described in the associated publication.