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(Plasmid #48696)


Item Catalog # Description Quantity Price (USD)
Plasmid 48696 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.


  • Vector backbone
  • Backbone manufacturer
  • Backbone size w/o insert (bp) 5400
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
  • Growth Strain(s)
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    8 copies of the amber suppressor tRNACUA-Leu
  • Species
  • Promoter human H1 promoter

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site BglII (not destroyed)
  • 3′ cloning site BglII (destroyed during cloning)
  • 5′ sequencing primer NA
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
  • Species
  • Mutation
    mutant E.coli leucyl synthetase specific for Anap (U35 and A36 are mutated to C and U, respectively)
  • Promoter CMV

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site XbaI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer BGH-Rev
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

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

Depositor Comments

The amber suppressor tRNACUA-Leu was derived from E. coli tRNAUAA-Leu in which U35 and A36 are mutated to C and U, respectively. The gene lacking 3′-CCA but with a 3′-TTTTTT sequence, driven by the human H1 promoter was synthesized by IDT. This was PCR amplified, digested with BglII and BamHI and was inserted into the BglII site of pCDNA3.1. Additional copies of the tRNA expression cassette were introduced in the resulting plasmid by successive rounds of BglII digestion, ligation of the BglII/BamHI digested insert, and verification of the insert orientation in the resulting plasmid by DNA sequencing and digestion analysis. AnapRS was PCR amplified and was inserted into pCDNA3.1-8x H1-tRNACUA-EcLeu between HindIII and XhoI restriction sites to generate pAnap.

Note that any small discrepancies between Addgene's quality control sequence and the assembled sequence from the depositor should not affect plasmid function.

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:

    pANAP was a gift from Peter Schultz (Addgene plasmid # 48696 ; ; RRID:Addgene_48696)
  • For your References section:

    A genetically encoded fluorescent probe in Mammalian cells. Chatterjee A, Guo J, Lee HS, Schultz PG. J Am Chem Soc. 2013 Aug 28;135(34):12540-3. doi: 10.1021/ja4059553. Epub 2013 Aug 14. 10.1021/ja4059553 PubMed 23924161