Addgene - pMYO127TAG-SepT Plasmid Data

Plasmid 34625: pMYO127TAG-SepT

Gene/insert name:
Myoglobin-127TAG
Insert size:
492
Species:
Sperm Whale: Physeter macrocephalus
Fusion protein or tag:
6xHIS
Terminal:
C terminal on insert
Mutation:
codon-optimized + an amber stop codon was introduced to the myoglobin gene at position Asp127
Vector backbone:
pSepT
(Search Vector Database)
Vector type:
Bacterial Expression
Modifications to Backbone:
To 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.
Promoter:
Ipp
Cloning site 5':
NotI
Site destroyed during cloning:
No
Cloning site 3':
BglII
Site destroyed during cloning:
No
5' sequencing primer:
pJR_F1 List of Sequencing Primers
3' sequencing primer:
M13 forward
Bacterial resistance(s)
Chloramphenicol
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
Low Copy
Sequence: View sequences (3)
Map:
View map
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Jesse Rinehart
Principal Investigator:
Dieter Soll
Terms and Licenses:
MTA

Comments:

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.

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

Article: Expanding the genetic code of Escherichia coli with phosphoserine. Park et al (Science. 2011 Aug 26;333(6046):1151-4. 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 34625" in your Materials and Methods section.