pUltra-dox
(Plasmid #58749)

• Purpose: (Empty Backbone) doxycycline inducible multicistronic lentiviral gene expression system.
• Depositing Labs: Yildirim Dogan, Kitai Kim
• Publication: pUltra plasmids (unpublished)

Backbone

• Vector backbone: pUltra-Chili
• Backbone manufacturer: Moore Lab, Addgene plasmid 48687
• Modifications to backbone: replace Ubic-promoter in pUltra-chili with Tre2. Tre2 was amplified from pSLIK-Neo-TGmiR-luc (Addgene plasmid 25745).
• Vector type: Mammalian Expression, Lentiviral, RNAi
• Promoter: TRE2 upstream of minimal CMV promoter

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: DsRed1-C

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid needs to be used with a second plasmid containing a constitutively expressed puromycin-selectable RTTA3 (Reverse Tet-Transactivator)--Addgene plasmids 58750 and 58751.

The advantage to other dox-inducible systems are:
1) better silencing in off condition because of the newer generation TRE2/RTTA3
2) you can induce to express two genes of interest (GOI) at the same time from one plasmid.
3) when the system is induced by dox, besides the GOI the tomato-reporter is induced to be expressed too, so you can actually see that the genes must be switched on.

loxP5171 at positions after cppt and before WPRE