Addgene - Flag pLJM1 RagD 77L Plasmid Data

Plasmid 19317: Flag pLJM1 RagD 77L

Gene/insert name:
Ras-related GTP binding D
Alt name:
RRAGD
Insert size:
1200
Species:
H. sapiens (human)
Entrez Gene:
RRAGD (RAGD, bA11D8.2.1)
Fusion protein or tag:
Flag
Terminal:
N terminal on insert
Mutation:
S77L
Vector backbone:
pLJM1
(Search Vector Database)
Vector type:
Mammalian Expression, Lentiviral
Backbone size w/o insert (bp):
7400
Cloning site 5':
SalI
Site destroyed during cloning:
No
Cloning site 3':
EcoRV
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 Exel10Gold or Stbl2 cells
High or low copy:
High Copy
Selectable markers:
Puromycin
Sequence: View sequences (2)
Principal Investigator:
David Sabatini
Terms and Licenses:
MTA

Comments:

pLJM1 is based on pLKO.1. pLJM1 was cut with AgeI and EcoRI and a Flag tag was cloned in. The cDNA was cloned into Flag-pLJM1 with SalI and EcoRV. The sequence across the MCS includes AgeI-Flag-SalI-gene-EcoRV-EcoRI

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

Article: The Rag GTPases bind raptor and mediate amino acid signaling to mTORC1. Sancak et al (Science. 2008 Jun 13. 320(5882):1496-501. 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 19317" in your Materials and Methods section.