|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||34625||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Modifications to backboneTo construct plasmid pSepT, the full-length gene encoding tRNA^Sep was constructed from overlapping oligonucleotides and ligated immediately downstream of the lpp promoter in pTECH (Bunjun et al., 2000) using EcoRI and BamHI restriction sites.
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
SpeciesSperm Whale: Physeter macrocephalus
Insert Size (bp)492
Mutationcodon-optimized + an amber stop codon was introduced to the myoglobin gene at position Asp127
- Promoter Ipp
/ Fusion Protein
- 6xHIS (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site BglII (not destroyed)
- 5′ sequencing primer pJR_F1
- 3′ sequencing primer M13 forward (Common Sequencing Primers)
To prevent possible enzymatic dephosphorylation of O-phospho-L-serine (Sep) in vivo, the gene encoding phosphoserine phosphatase (serB), which is catalyzing the last step in serine biosynthesis, was deleted from Escherichia coli strains Top10 (Top10∆serB - Addgene #34928) and BL21 (BL21∆serB - Addgene #34929). These strains are required hosts when using this plasmid.
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:pMYO127TAG-SepT was a gift from Jesse Rinehart & Dieter Söll (Addgene plasmid # 34625 ; http://n2t.net/addgene:34625 ; RRID:Addgene_34625)
For your References section:Expanding the genetic code of Escherichia coli with phosphoserine. Park HS, Hohn MJ, Umehara T, Guo LT, Osborne EM, Benner J, Noren CJ, Rinehart J, Soll D. Science. 2011 Aug 26;333(6046):1151-4. 10.1126/science.1207203 PubMed 21868676