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pMYO
(Plasmid #34626)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 34626 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pSepT
  • 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.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    Myoglobin
  • Species
    Sperm Whale: Physeter macrocephalus
  • Insert Size (bp)
    492
  • Mutation
    codon-optimized
  • Promoter lpp
  • Tag / Fusion Protein
    • 6xHIS (C terminal on insert)

Cloning Information

  • 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)

Resource Information

Depositor Comments

This plasmids is delivered in wild-type DH5alpha cells.

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.

How to cite this plasmid ( Back to top)

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:

    pMYO was a gift from Jesse Rinehart & Dieter Söll (Addgene plasmid # 34626 ; http://n2t.net/addgene:34626 ; RRID:Addgene_34626)
  • 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