EasyCloneYALI Collection
(Kit #
1000000140, 1000000141
)
Depositing Lab: Irina Borodina
The EasyCloneYALI collection contains two plasmid toolkits:
Kit #1000000140
The EasyCloneYALI- Integrative Vector Set contains integrative plasmids suitable for (over-)expression of (heterologous) genes in Y. lipolytica strains.
Kit #1000000141
The EasyCloneYALI- CRISPR/Cas9 Set contains plasmids for genome editing of Y. lipolytica via CRISPR/Cas9 technology.
Both kits are shipped as bacterial glycerol stocks in a 96-well plate format.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Kit | 1000000140 | EasyCloneYALI- Integrative Vector Set | 1 | $ 400 | Add to Cart | |
| Kit | 1000000141 | EasyCloneYALI- CRISPR/Cas9 Set | 1 | $ 400 | Add to Cart | |
Description
Kit #1000000140: EasyCloneYALI- Integrative Vector Set
The EasyCloneYALI- Integrative Vector Set contains a set of integrative vectors suitable for (over-)expression of (heterologous) genes in Y.lipolytica strains. The vectors allow for selection in prototrophic yeast strains (two different dominant selection markers and a uracil marker for auxothrophic strains). The EasyClone system provides the possibility for cloning of up to two genes when a bidirectional promoter (of any choice) is used, and integration of the construct into 11 well-defined 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 the possibility of repeated rounds of gene insertions.
Figure 1 legend: The collection contains integrative vectors with dominant and auxotrophic selection markers.
Kit #1000000141: EasyCloneYALI- CRISPR/Cas9 Set
The EasyCloneYALI- CRISPR/Cas9 Set contains plasmids for genome editing of Y. lipolytica via CRISPR/Cas9 technology and markerfree EasyClone vectors. The collection contains integrative Cas9 expression vectors and an episomal vector for gRNA expression. Additionally, the collection contains a set of markerfree EasyClone integration vectors and their corresponding gRNA vectors. The system can be used in Y. lipolytica strains, which are sensitive to the antibiotics nourseothricin and hygromycin, or can not use D-serine as a nitrogen source. The markerfree EasyClone vector system provides a possibility of cloning up to two genes when a bidirectional promoter (of any choice) is used, and integration of the construct into well-defined yeast chromosomal locations without the use of selection markers. Particular BioBricks are assembled and cloned into the vectors via Uracil-Specific Excision Reaction (USER) cloning.
Figure 2 legend: The collection contains plasmids for genome editing of Y. lipolytica via CRISPR/Cas9 technology and markerfree EasyClone vectors for (over-)expression of (heterologous) genes.
Depositor Information
User Manuals
User Manual: EASYCLONEYALI Integrative vector set for Y. lipolytica 1.2Â MBUser Manual: EASYCLONEYALI- CRIPSR/Cas9 genome editing of Y. lipolytica 1.5Â MB
Original Publication
EasyCloneYALI: CRISPR/Cas9-Based Synthetic Toolbox for Engineering of the Yeast Yarrowia lipolytica. Holkenbrink C, Dam MI, Kildegaard KR, Beder J, Dahlin J, Doménech Belda D, Borodina I.Biotechnol J.2018. Sep;13(9):e1700543. doi: 10.1002/biot.201700543. PubMed PMID 29377615
How to Cite these Kits
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 EasyCloneYALI- Integrative Vector Set was a gift from Irina Borodina (Addgene Kit #1000000140).”
“The EasyCloneYALI- CRISPR/Cas9 Set was a gift from Irina Borodina (Addgene Kit #1000000141).”
EasyCloneYALI- Integrative Vector Set - #1000000140
- 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 | pCfB4586 | Ampicillin |
| A / 2 | pCfB4778 | Ampicillin |
| A / 3 | pCfB5791 | Ampicillin |
| A / 4 | pCfB4132 | Ampicillin |
| A / 5 | pCfB5935 | Ampicillin |
| A / 6 | pCfB6577 | Ampicillin |
| A / 7 | pCfB6578 | Ampicillin |
| A / 8 | pCfB4796 | Ampicillin |
| A / 9 | pCfB6574 | Ampicillin |
| A / 10 | pCfB6575 | Ampicillin |
| A / 11 | pCfB6576 | Ampicillin |
| A / 12 | pCfB5219 | Ampicillin |
| B / 1 | pCfB6580 | Ampicillin |
| B / 2 | pCfB6581 | Ampicillin |
| B / 3 | pCfB4848 | Ampicillin |
| B / 4 | pCfB4788 | Ampicillin |
| B / 5 | pCfB4781 | Ampicillin |
| B / 6 | pCfB4782 | Ampicillin |
| B / 7 | pCfB4786 | Ampicillin |
| B / 8 | pCfB4780 | Ampicillin |
| B / 9 | pCfB4666 | Ampicillin |
| B / 10 | pCfB4779 | Ampicillin |
| B / 11 | pCfB4783 | Ampicillin |
| B / 12 | pCfB4787 | Ampicillin |
| C / 1 | pCfB4784 | Ampicillin |
| C / 2 | pCfB4785 | Ampicillin |
Kit Plate Map - #1000000140
EasyCloneYALI- CRISPR/Cas9 Set - #1000000141
- 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 | pCfB4906 | Ampicillin |
| A / 2 | pCfB6364 | Ampicillin |
| A / 3 | pCfB6682 | Ampicillin |
| A / 4 | pCfB6371 | Ampicillin |
| A / 5 | pCfB6684 | Ampicillin |
| A / 6 | pCfB6677 | Ampicillin |
| A / 7 | pCfB6681 | Ampicillin |
| A / 8 | pCfB6679 | Ampicillin |
| A / 9 | pCfB6627 | Ampicillin |
| A / 10 | pCfB6630 | Ampicillin |
| A / 11 | pCfB6631 | Ampicillin |
| A / 12 | pCfB6633 | Ampicillin |
| B / 1 | pCfB6637 | Ampicillin |
| B / 2 | pCfB6638 | Ampicillin |
| B / 3 | pCfB3405 | Ampicillin |