Addgene - pEnt L1L3 tTA-3 Plasmid Data

Plasmid 27105: pEnt L1L3 tTA-3

Gene/insert name:
tetracycline transactivator
Alt name:
tTA
Insert size:
1100
Vector backbone:
pEntr2B
(Search Vector Database)
Backbone manufacturer:
Invitrogen
Vector type:
Gateway entry vector
Backbone size w/o insert (bp):
2300
Cloning method:
Gateway Cloning
5' sequencing primer:
TCTACAAACTCTTCCTGTTAGT List of Sequencing Primers
3' sequencing primer:
AGAGATTTTGAGACACGGGC
Bacterial resistance(s)
Kanamycin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Sequence: View sequences (2)
Map:
View map
Principal Investigator:
Edward Hsiao
Terms and Licenses:
MTA
Tet Systems Limited Use Label License

Comments:

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Constitutive Gs activation using a single-construct tetracycline-inducible expression system in embryonic stem cells and mice Hsiao et al (Stem Cell Res Ther. 2011 Mar 4;2(2):11. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 27105" in your Materials and Methods section.