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pKT0232
(Plasmid #8727)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 8727 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pFA6a-link
  • Backbone size w/o insert (bp) 5444
  • Vector type
    Yeast Expression
  • Selectable markers
    KanR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    yECFP
  • Alt name
    Codon-optimized GFP variant
  • Alt name
    pKT232 (pFA6a–link–yECFP–13Myc–KANr)
  • Mutation
    yEGFP with F64L, S65T, Y66W, N146I, M153T, V163A
  • Tags / Fusion Proteins
    • Codon optimized linker (N terminal on backbone)
    • 13x Myc (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site PacI (not destroyed)
  • 3′ cloning site AscI (not destroyed)
  • 5′ sequencing primer SP6
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Full name of this plasmid is pFA6a-link-yECFP-13Myc-KanR. yECFP is created by mutagenesis from a codon-optimized green fluorescent protein - yEGFP1 (Cormack et al. 1997). This yEGFP variant is cloned into pDH5, replacing YFP. An codon-optimized linker was added into the yEGFP variant to improve expression level. The G418 resistance marker (KanR) was introduced into the plasmid by subcloning the BglII/EcoRI fragment of pDH3 (Hailey et al., 2002) into the plasmid, in place of SpHIS5.

The yeast GFP used to create this plasmid contained a point mutation M233I that is present in all GFP variants derived from this original yEGFP. The mutation had no effect on fluorescence (see associated publication for more details).

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:

    pKT0232 was a gift from Kurt Thorn (Addgene plasmid # 8727)
  • For your References section:

    Optimized cassettes for fluorescent protein tagging in Saccharomyces cerevisiae. Sheff MA, Thorn KS. Yeast 2004 Jun;21(8):661-70. 10.1002/yea.1130 PubMed 15197731