EasyClone 2.0 Yeast Toolkit
(Kit #
1000000073
)
Depositing Lab: Irina Borodina
EasyClone 2.0 is an expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains.
This kit will be shipped as bacterial glycerol stocks in 96-well plate format.
Original Publication
EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains. Stovicek V, Borja GM, Forster J, Borodina I. J Ind Microbiol Biotechnol. 2015 Nov;42(11):1519-31. doi: 10.1007/s10295-015-1684-8. Epub 2015 Sep 16. PubMed .
Description
This collection contains a set of 25 integrative vectors suitable for (over-) expression of (heterologous) genes in both laboratory and industrial S. cerevisiae strains. The vectors allow for selection in auxotrophic yeast strains (four different auxotrophic markers) as well as in prototrophic yeast strains (six different dominant selection markers). The EasyClone system provides a possibility of cloning of up to two genes, when a bidirectional promoter (of any of choice) is used and integration of the construct into well-defined 11 yeast chromosomal locations. Particular biobricks are assembled and cloned into the vectors via Uracil-Specific Excision Reaction (USER) cloning. The marker genes are flanked with loxP sites allowing for marker rescue and recycling, providing a possibility of repeated rounds of gene insertions.
Depositor-provided Files
Kit Manual: Borodina EasyClone 2.0 User Manual 1.2 MB
Additional References
- EasyClone 2.0 is described in (Stovicek et al., 2015a). [PMID: 26376869]
- EasyClone with detailed description of the cloning procedure (Jensen et al., 2014). [PMID: 24151867]
- Original paper on characterization of integration sites (Mikkelsen et al., 2012). [PMID: 22326477]
- USER cloning method
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The EasyClone 2.0 kit was a gift from Irina Borodina (Addgene kit # 1000000073 ).”
For your Reference section:
EasyClone 2.0: expanded toolkit of integrative vectors for stable gene expression in industrial Saccharomyces cerevisiae strains. Stovicek V, Borja GM, Forster J, Borodina I. J Ind Microbiol Biotechnol. 2015 Nov;42(11):1519-31. doi: 10.1007/s10295-015-1684-8. Epub 2015 Sep 16. PubMed .
EasyClone 2.0 Yeast Toolkit - #1000000073
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pCfB2188 | Ampicillin |
| A / 2 | pCfB2189 | Ampicillin |
| A / 3 | pCfB2226 | Ampicillin |
| A / 4 | pCfB2374 | Ampicillin |
| A / 5 | pCfB2190 | Ampicillin |
| A / 6 | pCfB2227 | Ampicillin |
| A / 7 | pCfB2229 | Ampicillin |
| A / 8 | pCfB2191 | Ampicillin |
| A / 9 | pCfB2192 | Ampicillin |
| A / 10 | pCfB2228 | Ampicillin |
| A / 11 | pCfB2336 | Ampicillin |
| A / 12 | pCfB2193 | Ampicillin |
| B / 1 | pCfB2513 | Ampicillin |
| B / 2 | pCfB2223 | Ampicillin |
| B / 3 | pCfB2194 | Ampicillin |
| B / 4 | pCfB2375 | Ampicillin |
| B / 5 | pCfB2224 | Ampicillin |
| B / 6 | pCfB2195 | Ampicillin |
| B / 7 | pCfB2196 | Ampicillin |
| B / 8 | pCfB2399 | Ampicillin |
| B / 9 | pCfB2197 | Ampicillin |
| B / 10 | pCfB2400 | Ampicillin |
| B / 11 | pCfB2225 | Ampicillin |
| B / 12 | pCfB2198 | Ampicillin |
| C / 1 | pCfB2337 | Ampicillin |