|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||105168||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerClontech = pTRE-T2
- Backbone size (bp) 4847
Vector typeMammalian Expression
- Promoter Tet/CMV
Growth in Bacteria
Growth instructionsMethylation deficient cells from NEB to alleviate methylation sensitive restriction sites like Xba I. However not recommended for long-term propagation and storage.
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer AGG CGT ATC ACG AGG CCC TTT CGT
- 3′ sequencing primer TAT TAC CGC CTT TGA GTG AGC TGA (Common Sequencing Primers)
Terms and Licenses
A synthetic DNA containing the IRES element was cloned into EcoRI/Xba I site of pTRE-T2-miR-Puromycin vector after removal of the mIR cassette. First digest parental pTRE-T2-miR-Puromycin vector with EcoRI and Xba I to remove the miR cassette. Then the IRES cassette was ligated back to the EcoRI site and Xba I site. The IRES cassette contains the unique EcoRI site followed by unique NotI and Mlu I sites that is followed by the start of the IRES sequence. This cloning site will allow insertion of the 1st cDNA. On the 3’ end of IRES there are unique Spe I and Xba I cloning sites for insertion of the second cDNA into the cassette.
Please note- to use the BsaI and XbaI sites for cloning, you must use unmethylated DNA. You will need to isolate the DNA from a methylation deficient bacterial strain.
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:pTRE-T2-∆mIR-IRES-Puromycin was a gift from Michael E. Wright (Addgene plasmid # 105168 ; http://n2t.net/addgene:105168 ; RRID:Addgene_105168)