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pEVOL-pAzF[TAG]
(Plasmid #164579)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 164579 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pEVOL
  • Backbone size w/o insert (bp) 4179
  • Total vector size (bp) 6104
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert 1

  • Gene/Insert name
    Mj pCNFRS[TAG] (aaRS1)
  • Alt name
    pAzFRS (1)
  • Species
    Methanocaldococcus jannaschii (engineered)
  • Insert Size (bp)
    924
  • Mutation
    Y32L L65V F108W Q109M D158G I159P
  • Promoter araBAD

Cloning Information for Gene/Insert 1

  • Cloning method Gibson Cloning
  • 5′ sequencing primer ATGCCATAGCATTTTTATCC
  • 3′ sequencing primer CTTCTGCGTTCTGATTTAATCTG
    • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    Mj pCNFRS[TAG] (aaRS2)
  • Alt name
    pAzFRS (2)
  • Species
    Methanocaldococcus jannaschii (engineered)
  • Insert Size (bp)
    924
  • Mutation
    Y32L L65V F108W Q109M D158G I159P
  • Promoter glnS

Cloning Information for Gene/Insert 2

  • Cloning method Gibson Cloning
  • 5′ sequencing primer TCCCGGTCATCAATCATCC
  • 3′ sequencing primer AACTCAATATATTGCAGAGATCATG
    • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    Mj tRNA[TAG]
  • Alt name
    Mj tRNA[CTA]
  • Species
    Methanocaldococcus jannaschii
  • Insert Size (bp)
    77
  • Promoter proK

Cloning Information for Gene/Insert 3

  • Cloning method Gibson Cloning
  • 5′ sequencing primer CCTGCAGGTAATTCCGCTTCGCAACATGTGAGCACCGG
  • 3′ sequencing primer AATAAATCCATGGCAAATTCGACCCTGAGCTGCTCGAGCATGC
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Machinery plasmid for incorporation of para-Azidophenylalanine (pAzF), para-cyanophenylalanine (pCNF), and para-ethynylphenylalanine (pENF) at TAG codons.

Please visit https://doi.org/10.1101/2021.04.12.439361 for bioRxiv preprint.

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:

    pEVOL-pAzF[TAG] was a gift from Ryan Mehl (Addgene plasmid # 164579 ; http://n2t.net/addgene:164579 ; RRID:Addgene_164579)

  • For your References section:

    Genetic Incorporation of Two Mutually Orthogonal Bioorthogonal Amino Acids That Enable Efficient Protein Dual-Labeling in Cells. Bednar RM, Jana S, Kuppa S, Franklin R, Beckman J, Antony E, Cooley RB, Mehl RA. ACS Chem Biol. 2021 Nov 19;16(11):2612-2622. doi: 10.1021/acschembio.1c00649. Epub 2021 Sep 30. 10.1021/acschembio.1c00649 PubMed 34590824
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