Addgene - pAAV-Ptet-RFP-shR-rtTA Plasmid Data

Plasmid 35625: pAAV-Ptet-RFP-shR-rtTA

Gene/insert name:
None
Vector backbone:
pAAV with pTRIPZ elements
(Search Vector Database)
Backbone manufacturer:
Stratagene and Open Biosystems
Vector type:
Mammalian Expression, RNAi ; Tet inducible, AAV
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.
Promoter:
Ptet
Bacterial resistance(s)
Ampicillin
Growth strain(s)
Stbl3
Growth temperature (℃):
30
High or low copy:
Unknown
Sequence: View sequences (3)
Map:
View map
Supplemental document:
pAAV-Ptet-RFP-shR-rtTA.pdf (application/pdf)
Principal Investigator:
Howard Gu
Terms and Licenses:
MTA
Tet Systems Limited Use Label License

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Fluorescence-based evaluation of shRNA efficacy. Naughton et al (Anal Biochem. 2011 Oct 1;417(1):162-4. Epub 2011 Jun 13. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 35625" in your Materials and Methods section.