pEnt R3L2 TetO(fl)-2
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||24416||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 2700
Vector typeEntry Vector
Growth in Bacteria
- Cloning method Restriction Enzyme
- 5′ sequencing primer pENTR-F (Common Sequencing Primers)
Terms and Licenses
Contains Gateway R3L2 sites. Full length (7 repeat) TetO. SbfI, FseI, AvrII, and SalI sites can be used to insert gene of interest, driven by TetO. Contains insulator sequence
Plasmid was constructed as follows: The pEntr2B entry vector was digested with AflII and XhoI to remove the attL1 and ccdb genes. Oligos
containing the attR3 site flanked by AflII on the 5' end and a polylinker (NotI-PacI-Bsu36I-NdeI-XhoI) on the 3' end were ligated to generate the empty pEntR3L2-MCS intermediate (Addgene plasmid # 26799). Using PCR cloning, the TetO-beta globin
intron sequence from pTetO (pUHD10.3) was cloned into the NotI/PacI sites in the reverse orientation,
and a 3' SbfI site was introduced. The pA sequence from pDest27 was then cloned into the NotI/SbfI sites, again in reverse orientation. A LoxP site was introduced into the NdeI/XhoI sites, allowing full
excision of the targeted construct when used together with the LoxP site in the pEntl1L3 vector. The CBG core
insulator was then cloned into the Bsu36I/NdeI sites to generate the starting entry vector pEntR3L2 TetO(fl)-2.
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:pEnt R3L2 TetO(fl)-2 was a gift from Edward Hsiao (Addgene plasmid # 24416 ; http://n2t.net/addgene:24416 ; RRID:Addgene_24416)
For your References section:Constitutive Gs activation using a single-construct tetracycline-inducible expression system in embryonic stem cells and mice. Hsiao EC, Nguyen TD, Ng JK, Scott MJ, Chang WC, Zahed H, Conklin BR. Stem Cell Res Ther. 2011 Mar 4. 2(2):11. 10.1186/scrt52 PubMed 21375737