pLenti CMV rtTA3 Blast (w756-1)
Purpose3rd gen lentiviral reverse tetracycline-controlled transactivator 3 (rtTA3) expression vector, CMV promoter, Blast
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26429||Plasmid sent as bacteria in agar stab||1||$65|
Virus (1 mL at titer > 5x10⁵ TU/mL)
and Plasmid. More Information
This material is available to academics and nonprofits only.Viral services are available to scientists in the United States and select countries. Additional information
Vector backbonep156RRL-sinPPT-CMV-GFP-PRE/Nhe I
- Backbone size w/o insert (bp) 7681
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Growth instructionsStbl3, 37oC.
Copy numberHigh Copy
Gene/Insert nameTetracycline repressor A3 mutant
Insert Size (bp)707
- Cloning method Restriction Enzyme
- 5′ sequencing primer CMVforw
- 3′ sequencing primer WPRErev (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byThe rtTA3 element was a gift from Dr. Dominic Esposito.
Terms and Licenses
Articles Citing this Plasmid
Use to make them Tet-On Advanced.
There are a few mismatches between Addgene's sequence and Dr Campeau's sequence. The sequence difference does not affect expression of the rtTA3.
Information for Lentiviral Prep (Catalog # 26429-LV) ( Back to top )
Ready-to-use Lentiviral Prep particles produced from pLenti CMV rtTA3 Blast (w756-1) (#26429). In addition to the viral particles, you will also receive purified pLenti CMV rtTA3 Blast (w756-1) plasmid DNA.
Lentiviral particles carrying the rtTA3 transactivator protein and blasticidin resistance.
- Volume 1 mL
- Titer ≥5x10⁵ TU/mL
- Pricing $220 USD for preparation of 1 mL virus + $30 USD for plasmid.
- Storage Store at -80℃. Thaw just before use and keep on ice.
- Shipment Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
Viral Production & Use
Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide
Viral Quality Control
- Colony formation assay: A549 cells were transduced with serial dilutions of 26429-LV and treated with blasticidin. Blasticidin-resistant colonies were expanded for approximately 2 weeks, stained with crystal violet, and counted.
- PCR confirmation of insert: PCR was carried out with primers targeting the CMV promoter and WPRE. The PCR product was visualized on an agarose gel for size confirmation.
Forward Primer: CMV-F GTAACAACTCCGCCCCATT
Reverse Primer: WPRE-R CATAGCGTAAAAGGAGCAACA
- Confirmation of protein expression: A549 cells were transduced with 26429-LV at an MOI of 1 and treated with blasticidin. Polyclonal pools of blasticidin-resistant cells were expanded, collected, lysed and tested for rtTA expression via immunoblotting. You can view the stable cell line expression data here or in the image section at the top of this page. Read our protocol for generating stable cell lines here.
Visit our viral production page for more information.
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:pLenti CMV rtTA3 Blast (w756-1) was a gift from Eric Campeau (Addgene plasmid # 26429)
Generated by Addgene from full sequence.
Map uploaded by Addgene staff.
Map uploaded by the depositor.