Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||17655||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3000
Vector typeBacterial Expression
Growth in Bacteria
Growth instructions30 degrees, Stbl3
Copy numberHigh Copy
Gene/Insert namelac I-SceI tet
Alt namelac operator
Alt nametet operator
- Cloning method Restriction Enzyme
- 5′ sequencing primer T7
- 3′ sequencing primer T3 (Common Sequencing Primers)
The 256 lac operator repeats (XhoI fragment) and the 96 tet operator repeats (XhoI –SacII fragment) from the p16PCbeta
plasmid (gift from D. Spector) were cloned into pBluescript. Between the two fragments a single 18nt I-SceI site was inserted (SalI).
lac operator repeat: tggaattgtgagcggataacaatt
tet operator repeat: tccctatcagtgatagaga
This plasmid is not stable in bacteria. Every bacterial stock made will contain correct and incorrect bacteria and when bacteria are grown, you will get colonies which contain the correct plasmid and others which contain recombinant versions. If you check multiple colonies (10-20) in the samples after streaking out a stab, you should find correct ones. The recipient will need to grow the bacteria, isolate individual colonies check them, and make a prep from the correct one. We recommend processing the stab and screening colonies immediately upon receipt.
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:Lac-I-SceI-Tet was a gift from Tom Misteli (Addgene plasmid # 17655 ; http://n2t.net/addgene:17655 ; RRID:Addgene_17655)
For your References section:Positional stability of single double-strand breaks in mammalian cells. Soutoglou E, Dorn JF, Sengupta K, Jasin M, Nussenzweig A, Ried T, Danuser G, Misteli T. Nat Cell Biol. 2007 Jun . 9(6):675-82. 10.1038/ncb1591 PubMed 17486118