PurposeEncodes Thr251Gly-EcPheRS Under IPTG-Inducible (PT5) Control
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||62598||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) 4541
- Total vector size (bp) 7897
Modifications to backboneThe genes encoding wild-type E. coli PheRS were isolated by genomic DNA extraction from E. coli DH10B and PCR amplification. The purified fragments were ligated into pQE-80L-Kan (SphI, HindIII). The resulting plasmid was subjected to site-directed mutagenesis to generate the Thr251Gly mutation.
Vector typeBacterial Expression
Growth in Bacteria
Copy numberHigh Copy
Alt nameE. coli Phenylalanyl-tRNA Synthetase (Thr251Gly)
Insert Size (bp)3386
- Promoter T5
/ Fusion Protein
- 6xHis (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site SphI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer pQE Promoter
- 3′ sequencing primer pQE Reverse (Common Sequencing Primers)
Please note that this plasmid may require a unique bacterial strain, so make sure to confirm that you can also obtain the appropriate growth strain. Please contact us at [email protected] if you have any questions.
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:pKPY514 was a gift from David Tirrell (Addgene plasmid # 62598 ; http://n2t.net/addgene:62598 ; RRID:Addgene_62598)
For your References section:Cell-specific proteomic analysis in Caenorhabditis elegans. Yuet KP, Doma MK, Ngo JT, Sweredoski MJ, Graham RL, Moradian A, Hess S, Schuman EM, Sternberg PW, Tirrell DA. Proc Natl Acad Sci U S A. 2015 Feb 17. pii: 201421567. 10.1073/pnas.1421567112 PubMed 25691744