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pcDNA3.1(+)_4x(U6 tRNA M15)_CMV NESPylRS(AF)
(Plasmid #182652)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 182652 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pcDNA3.1(+)
  • Vector type
    Mammalian Expression
  • Selectable markers
    Zeocin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    codon-optimized Y306A/Y384F (AF) double mutant of Methanosarcina mazei-derived pyrrolysine (Pyl) tRNA synthetase
  • Alt name
    NESPylRS(AF)
  • Species
    Methanosarcina mazei
  • Mutation
    Y306A/Y384F (AF) double mutant of Methanosarcina mazei-derived pyrrolysine (Pyl) tRNA synthetase
  • Promoter CMV
  • Tag / Fusion Protein
    • nuclear export signal (NES) (N terminal on insert)

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    4xU6-tRNAM15
  • Species
    Synthetic; Methanosarcina mazei
  • Mutation
    synthetic M15 tRNA(amber) variant
  • Promoter U6

Cloning Information for Gene/Insert 2

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    The codon-optimized gene of PylRS(AF) was custom synthesized by GeneScript. 4xU6-M15 tRNA cassette is derived from the following plasmid https://www.addgene.org/105830/

Terms and Licenses

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

Depositor Comments

Addgene QC NGS analysis identified 3X U6 tRNA M15, rather than 4X. The depositor has confirmed that this should have no influence on the functionality of the 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:

    pcDNA3.1(+)_4x(U6 tRNA M15)_CMV NESPylRS(AF) was a gift from Ivana Nikić-Spiegel (Addgene plasmid # 182652)

  • For your References section:

    Minimal genetically encoded tags for fluorescent protein labeling in living neurons. Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I. Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y. 10.1038/s41467-022-27956-y PubMed 35031604
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