Addgene - pLJM1 Flag Raptor HRas25 Plasmid Data

Plasmid 26637: pLJM1 Flag Raptor HRas25

Gene/insert name:
raptor-HRas25
Alt name:
raptor HRas25
Species:
H. sapiens (human)
Entrez Gene:
RPTOR (KOG1, Mip1)
Fusion protein or tag:
Flag
Terminal:
N terminal on insert
Vector backbone:
pLJM1
(Search Vector Database)
Backbone manufacturer:
Available at Addgene (#19319)
Vector type:
Mammalian Expression, Lentiviral
Backbone size w/o insert (bp):
6600
Cloning site 5':
AgeI
Site destroyed during cloning:
No
Cloning site 3':
BstBI
Site destroyed during cloning:
No
5' sequencing primer:
CMV-F List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
Stbl3
Growth temperature (℃):
37
Growth instructions:
Grow in XL10Gold or Stbl3 cells to decrease recombination. Growing the cultures at 30C rather than 37 and in SOC sometimes helps with yield.
High or low copy:
Unknown
Selectable markers:
Puromycin
Sequence: View sequences (2)
Map:
View map
Principal Investigator:
David Sabatini
Terms and Licenses:
MTA

Comments:

The insert is structured:

AgeI-FlagTag-SalI-Raptor without stop codon-EcoRV-localization tag-stop codon-BstBI.

New genes can be cloned into the vector with N-term Flag and C-term localization
signal using SalI-EcoRV.

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

Article: Ragulator-Rag complex targets mTORC1 to the lysosomal surface and is necessary for its activation by amino acids. Sancak et al (Cell. 2010 Apr 16. 141(2):290-303. 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 26637" in your Materials and Methods section.