pINDUCER21 (ORF-EG)
(Plasmid #46948)

• Purpose: (Empty Backbone) Inducible lentiviral gene expression vector. Chloramphenicol cm + ccdB cassette may be removed to clone in gene of interest
• Depositing Labs: Stephen Elledge, Thomas Westbrook
• Publication: Meerbrey et al Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 10.1073/pnas.1019736108. Epub 2011 Feb 9.

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, 25 & 100 μg/mL
• 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

Resource Information

• Supplemental Documents:
  • Viral Production Protocol
• Articles Citing this Plasmid:
  • 25 References

Terms and Licenses

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

Information for Cloning Grade DNA (Catalog # 46948-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