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pUltra-hot
(Plasmid #24130)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 24130 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

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
    • mCherry (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
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

pUltra was cloned by Yildirim Dogan ([email protected])

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

By cloning into the compatible cloning sites* (XbaI and BamHI) downstream of mCherry-P2A, you get a bi-cistronic expression of mCherry and the gene of interest. You can clone a second gene of interest into the NheI/bclI**donwstream of mCherry-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: 3971-4033 and T2A is between bp 4049-4111

How to cite this plasmid ( Back to top)

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:

    pUltra-hot was a gift from Malcolm Moore (Addgene plasmid # 24130 ; http://n2t.net/addgene:24130 ; RRID:Addgene_24130)