FU-tetO-Gateway
(Plasmid #43914)

• Purpose: (Empty Backbone)
• Depositing Lab: John Gearhart
• Publication: Addis et al PLoS One. 2011;6(12):e28719. doi: 10.1371/journal.pone.0028719. Epub 2011 Dec 9.

Backbone

• Vector backbone: FU-tet-o-hOct4
• Backbone manufacturer: Addgene Plasmid 19778
• Backbone size (bp): 8600
• Modifications to backbone: OCT4 ORF replaced with Gateway recombination cassette (from pEF-DEST51, Invitrogen)
• Vector type: Mammalian Expression, Lentiviral ; Gateway Destination vector, Doxycycline inducible
• Promoter: TRE promoter, Tet-ON
• Selectable markers: Zeo marker is outside the LTRs and will not be packaged into virus.
• Tags / Fusion Proteins:
  • V5 (C terminal on backbone)
  • 6xHis (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: Use ccdb-resistant cells, such as OneShot Survival ccdb Competent Cells (Invitrogen)
• Copy number: Low Copy

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: LNCX; T7
• 3′ sequencing primer: WPRE-R

Resource Information

• Supplemental Documents:
  • FU-tetO-Gateway annotated genbank file
• A portion of this plasmid was derived from a plasmid made by: Backbone plasmid FU-tetO-hOCT4 from Addgene (Plasmid #19778, Konrad Hochedlinger lab); Gateway cassette from pEF-DEST51 from Invitrogen.
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

To make a drug-inducible lentiviral vector that is compatible with Gateway recombination (Invitrogen), the depositing laboratory removed the OCT4 open reading frame from FU-tetO-hOCT4 (Addgene plasmid 19778) and replaced it with a Gateway cassette cloned from pEF-DEST51 (Invitrogen) to generate FU-tetO-Gateway. ORFs that lack stop codons can be cloned into this plasmid to express proteins with a C-terminal V5 epitope tag.

Please note that Addgene's sequencing results identified a single nucleotide insert between bp# 2913/2914 and single nucleotide mismatches at bp# 3303 and 4652 when compared to the full plasmid sequence provided by the depositing laboratory. These differences do not affect the function of the plasmid.