This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.
We have implemented updates to our Transparency and Privacy Policy in light of the new GDPR standards. When logging in or creating a new account, you will be asked to read and acknowledge these policy changes. Additionally, you can find our Transparency and Privacy Policy here.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

pINDUCER21 (ORF-EG)
(Plasmid #46948)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 46948 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pHAGE-CAGS-W
  • Backbone manufacturer
    Dr. Richard Mulligan
  • Backbone size (bp) 4874
  • Modifications to backbone
    pINDUCER21 (ORF-UG) was made by digesting pINDUCER20 with NdeI and SphI. eGFP was PCR amplified with AgeI and SphI on the 5′ and 3′ ends, respectively, and then digested with AgeI and SphI. The vector and eGFP were ligated and sequence verified. pINDUCER20 (ORF-UN) was made by digesting TRE2 out of the pSLIK-Neo- TGmiR-Luc vector with BamHI and SpeI and blunt-end cloned into the backbone of SpeI and XhoI digested pHAGE-CAGS-W vector (Dr. Richard Mulligan, Harvard Institute of Medicine, Boston) to generate the pHAGE-TRE vector. A fragment containing the gateway recipient cassette was cut out of the pHAGE-EF-DEST vector (G.H.) with SpeI and inserted into pHAGE-TRE to generate the pHAGE-TRE-DEST vector. The Ubc-rtTA3-IRES-Neo fragment was cut out of the pSLIK-Neo-TGmiR-Luc vector with PacI and ClaI and bluntend cloned into the backbone of SphI and ClaI digested pHAGE-TRE-DEST.
  • Vector type
    Lentiviral
  • Selectable markers
    eGFP

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol and Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    ccdB Survival
  • Growth instructions
    pInducer 21 should be grown in LB media containing 50ug/ml ampicillin + 30ug/ml Chloramphenicol. Easiest to grow this gene expression vector at 30 deg C. When transforming pInducer 21 (parent vector alone) into ecoli, use competant ecoli strain "ccdB" Invitrogen cat# A10460. Following replacement of ccdB surival + Chloramphenicol cassette w cDNA of interest, resultant plasmid is then ampicillin resistant only, and can be transformed into standard competant ecoli strains such as DH5alpha, Stbl3(minimizes LTR recombination), or NEB10
  • Copy number
    Unknown
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:

    pINDUCER21 (ORF-EG) was a gift from Stephen Elledge & Thomas Westbrook (Addgene plasmid # 46948)
  • For your References section:

    The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo. Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF, Elledge SJ. Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 10.1073/pnas.1019736108. Epub 2011 Feb 9. 10.1073/pnas.1019736108 PubMed 21307310