pEnt L1L3 tTA-3
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||27105||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 2300
Vector typeGateway entry vector
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nametetracycline transactivator
Insert Size (bp)1100
- Cloning method Gateway Cloning
- 5′ sequencing primer TCTACAAACTCTTCCTGTTAGT
- 3′ sequencing primer AGAGATTTTGAGACACGGGC (Common Sequencing Primers)
Gateway L1L3 entry vector containing the tTA regulator. Promoters can be inserted using PacI and SbfI. Does not contain insulator sequence.
Plasmid was constructed as follows: The pEntr2B entry vector (Invitrogen) was digested with PstI and XhoI to remove the attL2 site.
Oligonucleotides containing SalI-XhoI-Bsu36I-NdeI-PacI-EcoRI-NotI flanking the attL3 sequence on the 5' side
and PstI on the 3' side were ligated in to create an intermediate plasmid carrying attL1 and attL3 sites. A LoxP sequence was introduced at the XhoI site. The tTA and pA sequences were subsequently cloned into the EcoRI/NotI sites by PCR cloning from pUHG15-1 to generate pEntL1L3 tTA-3.
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 L1L3 tTA-3 was a gift from Edward Hsiao (Addgene plasmid # 27105 ; http://n2t.net/addgene:27105 ; RRID:Addgene_27105)
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