pAS1NB m Rosella I
Purposea biosensor comprised of a fast-maturing pH-stable red fluorescent protein fused to a pH-sensitive green fluorescent protein variant. Mitochodrial localization.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||71247||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Total vector size (bp) 9209
Vector typeYeast Expression
Growth in Bacteria
Alt namesuper ecliptic pHluorin (SEP)
- Promoter PGK1
/ Fusion Protein
- yeast mitochondrial targeting sequence of citrate synthase (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer PPGK1-F (cagcctgttctcacacactc)
- 3′ sequencing primer yPGK1-term-R (AGCGTAAAGGATGGGGAAAG) (Common Sequencing Primers)
The primer pair CIT2Up (5'‑GAAGATCTAAACAATGTCAG TCGATATTATC) and CIT2Do (5'‑GAGGATCCGTAATTTCA GCAAATCTCTCC) was used to amplify the yeast mitochondrial targeting sequence of citrate synthase flanked by BglII sites from yeast genomic DNA. The resultant PCR fragment, following digestion with BglII, was ligated in‑frame at the 5' end of the Rosella expression cassette.
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:pAS1NB m Rosella I was a gift from Mark Prescott (Addgene plasmid # 71247 ; http://n2t.net/addgene:71247 ; RRID:Addgene_71247)
For your References section:Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in yeast. Rosado CJ, Mijaljica D, Hatzinisiriou I, Prescott M, Devenish RJ. Autophagy. 2008 Feb;4(2):205-13. Epub 2007 Nov 21. 5331 [pii] PubMed 18094608