|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||8736||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5014
Vector typeYeast Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Alt nameCodon-optimized GFP variant
Alt namepKT220 (pFA6a–link–yECitrine–3HA–KANr)
MutationyEGFP with S65G, V68L, Q69M, S72A, T203Y
/ Fusion Proteins
- Codon optimized linker (N terminal on backbone)
- 3x HA (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site PacI (not destroyed)
- 3′ cloning site AscI (not destroyed)
- 5′ sequencing primer SP6 (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Full name of this plasmid is pFA6a-link-yECitrine-3HA-KanR. yECitrine 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).
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:pKT0220 was a gift from Kurt Thorn (Addgene plasmid # 8736 ; http://n2t.net/addgene:8736 ; RRID:Addgene_8736)
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