pUltra
(Plasmid #24129)

• Purpose: (Empty Backbone) 3rd generation Lentiviral vector for bi-cistronic expression of EGFP and the gene of interest
• Depositing Lab: Malcolm Moore
• Publication: Lou et al PLoS One. 2012;7(3):e33093. doi: 10.1371/journal.pone.0033093. Epub 2012 Mar 9.

Backbone

• Vector backbone: pRRL
• Backbone manufacturer: Didier Trono
• Backbone size (bp): 7000
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Stbl3 or SC110 (See below**)
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: None
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI/NheI (not destroyed)
• 3′ cloning site: BamHI/BcII (not destroyed)
• 5′ sequencing primer: hUBCpro-F

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 61 References

Terms and Licenses

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

Depositor Comments

pUltra was cloned by Yildirim Dogan () It is described in the associated article (PMID 22427958) and also in PMID 24630793.

This is a 3rd generation Lentiviral vector with an internal Ubiquitinc Promoter.

By cloning into the compatible cloning sites* (XbaI and BamHI) downstream of EGFP-P2A, you get a bi-cistronic expression of EGFP and the gene of interest. You can clone a second gene of interest into the NheI/bclI**donwstream of EGFP-P2A-gene1-T2A and get a multi-cistronic expression of all three genes.

* compatible cloning site (ccs): you can pcr amplifiy your gene of interest with a forward primer including one of these cutting sites: SpeI or NheI or XbaI and a reverse primer with one these cutting sites: BglII or BamHI or bclI. The PCR product is now compatible with the first site as well the 2nd site. "You can mix and match".

** for bclI the vector has to be amplified by dam-methylation defective E coli strains (e.g. SC110 Stratagene).

Further advantage of the vector is, that you can simultaneously do RNAi by cloning H1-shRNA cassettes into the unique SnaBI site in the 3´-LTR. Plus while integration the RNAi cassette gets doubled, since the 5´LTR is dublicated while reverse transcripton from the 3´-LTR

***P2A: 3984-4046 bp and T2A: 4062-4124 bp

Information for Cloning Grade DNA (Catalog # 24129-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

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

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here