|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||43914||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene Plasmid 19778
- Backbone size (bp) 8600
Modifications to backboneOCT4 ORF replaced with Gateway recombination cassette (from pEF-DEST51, Invitrogen)
Vector typeMammalian Expression, Lentiviral ; Gateway Destination vector, Doxycycline inducible
- Promoter TRE promoter, Tet-ON
Selectable markersZeo marker is outside the LTRs and will not be packaged into virus.
/ 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 Strain(s)ccdB Survival
Growth instructionsUse ccdb-resistant cells, such as OneShot Survival ccdb Competent Cells (Invitrogen)
Copy numberLow Copy
- Cloning method Gateway Cloning
- 5′ sequencing primer LNCX; T7
- 3′ sequencing primer WPRE-R (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
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:FU-tetO-Gateway was a gift from John Gearhart (Addgene plasmid # 43914 ; http://n2t.net/addgene:43914 ; RRID:Addgene_43914)
For your References section:Efficient conversion of astrocytes to functional midbrain dopaminergic neurons using a single polycistronic vector. Addis RC, Hsu FC, Wright RL, Dichter MA, Coulter DA, Gearhart JD. PLoS One. 2011;6(12):e28719. doi: 10.1371/journal.pone.0028719. Epub 2011 Dec 9. 10.1371/journal.pone.0028719 PubMed 22174877