|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||33356||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene (#33361)
Vector typeMouse Targeting
Selectable markersNeomycin (select with G418) ; tetracycline
Growth in Bacteria
Growth instructionsDo not select for this plasmid with tetracycline, as we have found that the Tet resistance cassette does not work reliably in high-copy plasmids such as this one. The Tet cassette MUST be transferred to a low-copy vector, e.g. a BAC, to allow Tet resistance in a reliable fashion. In BAC clones it has been extremely reliable.
Gene/Insert nameEGFP-IRES-Bgeo + TetR
- Cloning method Restriction Enzyme
- 5′ cloning site none (unknown if destroyed)
- 3′ cloning site none (unknown if destroyed)
- 5′ sequencing primer EGFP-C
- 3′ sequencing primer LacZ-R (Common Sequencing Primers)
Terms and Licenses
To generate the GFP-IRES-β-geo cassette, a PacI site was inserted was inserted upstream of the Kozak consensus of EGFP in pEGFP-C1 (kind gift of David Piston) and a PacI/XhoI fragment containing the EGFP open reading frame was inserted into pIBG-FTET (Chandler et al., 2007b) to create pGFP-IBGFTet (also known as pGIBGFTET).
A mismatch between author sequence and Addgene sequencing results was found at nucleotide position 5695 (with SV40pA-R primer) as well as at position 6913 (with EBV-rev primer). These sequence differences predate the construction of this 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:pGFP-IBGFTet was a gift from Doug Mortlock (Addgene plasmid # 33356 ; http://n2t.net/addgene:33356 ; RRID:Addgene_33356)
For your References section:Identification of an ancient Bmp4 mesoderm enhancer located 46 kb from the promoter. Chandler KJ, Chandler RL, Mortlock DP. Dev Biol. 2009 Mar 15;327(2):590-602. Epub 2009 Jan 3. 10.1016/j.ydbio.2008.12.033 PubMed 19159624