Addgene - pcDNA3 Flag TSC2 Plasmid Data

Plasmid 14129: pcDNA3 Flag TSC2

Gene/insert name:
TSC2
Alt name:
tuberin
Insert size:
5400
Species:
H. sapiens (human)
Entrez Gene:
TSC2 (LAM, TSC4)
Fusion protein or tag:
Flag
Terminal:
N terminal on backbone
Vector backbone:
pcDNA3-Flag
(Search Vector Database)
Backbone manufacturer:
FLAG (MDYKDDDDK) was cloned between HindIII and BamHI of pcDNA3
Vector type:
Mammalian Expression
Backbone size w/o insert (bp):
5500
Cloning site 5':
BamHI
Site destroyed during cloning:
No
Cloning site 3':
XbaI
Site destroyed during cloning:
No
5' sequencing primer:
T7 List of Sequencing Primers
3' sequencing primer:
SP6
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
Neomycin
Person or lab that originally
cloned the gene/insert:
David J. Kwiatkowski
Sequence: View sequences (3)
Principal Investigator:
Brendan Manning
Terms and Licenses:
MTA

Comments:

The first 276 base pairs of TSC2 were PCR-amplified from pcDNA6-TSC2 and inserted between the BamHI and NotI sites in frame with the FLAG-coding sequence in pcDNA3-FLAG. The remainder of TSC2 was subcloned from pcDNA6-TSC2 and inserted between the NotI and XbaI sites of the above construct.

Diagnostic digests reveal an extra BamHI site in this plasmid. BamHI will not linearize the construct.

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

Article: Identification of the tuberous sclerosis complex-2 tumor suppressor gene product tuberin as a target of the phosphoinositide 3-kinase/akt pathway. Manning et al (Mol Cell. 2002 Jul . 10(1):151-62. 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 14129" in your Materials and Methods section.