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Plasmid kit icon

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 $330 Add to Cart
Kit 1000000141 EasyCloneYALI- CRISPR/Cas9 Set 1 $330 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.


EasyClone Integration Vector
  • 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.


EasyClone CRISPR/Cas9
  • 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

Icon User Manual: EASYCLONEYALI Integrative vector set for Y. lipolytica (1.2 MB)
Icon User Manual: EASYCLONEYALI- CRIPSR/Cas9 genome editing of Y. lipolytica (1.5 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
Data calculated @ 2019-10-14

Kit Plate Map - #1000000140

Plate #1

  1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H

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
Data calculated @ 2019-10-14

Kit Plate Map - #1000000141

Plate #1

  1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
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