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URL: http://www.addgene.org/74634
Proper Citation: RRID:Addgene_74634
Insert Name: mCherry
Bacterial Resistance: Ampicillin
Defining Citation: PMID:26994102
Vector Backbone Description: Backbone Marker:Janke et al., (2004) Yeast 21:947-962; Backbone Size:4940; Vector Backbone:pYM27; Vector Types:; Bacterial Resistance:Ampicillin
Comments: During verification of the constructed cassettes by restriction analysis and sequencing we found out that there are 254 bp missing after the BamHI site in between the fluorophore and the Ashbya gossypii TEF promotor region regulating expression of the selectable marker in pFM571, pIM572 and pIM573. We checked the original pYM27 plasmid (Janke, et al., 2004, Yeast 21:947-962 ) we had in our hands and compared the obtained sequence to the published one. We also checked by restriction analysis pYM27 from an independent source (but also obtained from Euroscarf) and got the same results. In the published pYM27 sequence, there is a sequence AGATCC resembling BamHI site (GGATCC) at position 1044-1049 bp. Likely due to a BamHI star activity that led to cutting this BamHI-like site in parallel with the true BamHI site, the nucleotides between these two sites got lost during construction of pYM27 or its predecessors and are missing in pYM27 and thus also in our pYM27-based modules - pFM571, pIM572 and pIM573. It seems, however, that the functionality of the vector and of the selection marker is not disturbed since transformants were readily obtained with all the concerned cassettes when selecting on geneticin.
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org
