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MoClo-YTK
(Kit # 1000000061 )

Depositing Lab:   John Dueber

This collection contains a set of 96 standardized and characterized parts that can be used for bottom-up hierarchical assembly of single and multi-gene constructs for S. cerevisiae expression. This kit adapts the MoClo stategy for assembly (but may not be compatible with other MoClo kits), and includes an assortment of parts (promoters, terminators, peptide tags, selection cassettes, etc).

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly. Lee ME, DeLoache, WC A, Cervantes B, Dueber, JE. ACS Synthetic Biology 2015 DOI: 10.1021/sb500366v. Journal .

Description

This collection contains a set of 96 standardized and characterized parts that can be used for bottom-up hierarchical assembly of single and multi-gene constructs for S. cerevisiae expression. The toolkit was deposited by John Dueber’s lab and contains characterized promoters, terminators, peptide tags, copy number machinery, and genome-editing tools. The hierarchical assembly scheme is based on the Golden Gate cloning technique, but it is also designed to be compatible with recombination based cloning such as yeast in vivo assembly, ligation independent cloning (SLIC) or Gibson Assembly. The design specifications of this collection also make constructs BioBrick-compatible. Use of this workflow allows for construction of multi-gene plasmids from PCR templates in three days.


Schematic of the Dueber Lab YTK system

Kit Documentation

Note from the depositor: We have noticed that some of the constructs are mildly toxic to E.coli (particularly those expressing fluorescent proteins), and will disappear from the population if a culture is passaged multiple times. The wells where we have noticed this issue are the following: A1, D11, E2, G11, G12, H1, H5, H6, H7, H11, H12. We suggest these be streaked onto solid media to pick fresh colonies every time. Conveniently, the fluorescent proteins themselves make it very easy to identify correct clones. Wells A1, D11, E2, H11, and H12 are green, while G11, G12, H1, H5, H6, H7 are red.


Documents describing this kit: Icon YTK summary (128.0 KB)


Download a ZIP file containing all MoClo-YTK plasmid sequences in GenBank format: Icon YTK Genbank files (256.1 KB)

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 MoClo-YTK plasmid kit was a gift from John Dueber (Addgene kit # 1000000061).”

For your Reference section:

A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly. Lee ME, DeLoache, WC A, Cervantes B, Dueber, JE. ACS Synthetic Biology 2015. DOI: 10.1021/sb500366v. Journal .

MoClo Yeast Toolkit - #1000000061

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 links to its kit plate map on the right. 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

Chloramphenicol
Ampicillin
Kanamycin
Spectinomycin

Inventory

Well Plasmid Resistance
A / 1 pYTK001
Chloramphenicol
A / 2 pYTK002
Chloramphenicol
A / 3 pYTK003
Chloramphenicol
A / 4 pYTK004
Chloramphenicol
A / 5 pYTK005
Chloramphenicol
A / 6 pYTK006
Chloramphenicol
A / 7 pYTK007
Chloramphenicol
A / 8 pYTK008
Chloramphenicol
A / 9 pYTK009
Chloramphenicol
A / 10 pYTK010
Chloramphenicol
A / 11 pYTK011
Chloramphenicol
A / 12 pYTK012
Chloramphenicol
B / 1 pYTK013
Chloramphenicol
B / 2 pYTK014
Chloramphenicol
B / 3 pYTK015
Chloramphenicol
B / 4 pYTK016
Chloramphenicol
B / 5 pYTK017
Chloramphenicol
B / 6 pYTK018
Chloramphenicol
B / 7 pYTK019
Chloramphenicol
B / 8 pYTK020
Chloramphenicol
B / 9 pYTK021
Chloramphenicol
B / 10 pYTK022
Chloramphenicol
B / 11 pYTK023
Chloramphenicol
B / 12 pYTK024
Chloramphenicol
C / 1 pYTK025
Chloramphenicol
C / 2 pYTK026
Chloramphenicol
C / 3 pYTK027
Chloramphenicol
C / 4 pYTK028
Chloramphenicol
C / 5 pYTK029
Chloramphenicol
C / 6 pYTK030
Chloramphenicol
C / 7 pYTK031
Chloramphenicol
C / 8 pYTK032
Chloramphenicol
C / 9 pYTK033
Chloramphenicol
C / 10 pYTK034
Chloramphenicol
C / 11 pYTK035
Chloramphenicol
C / 12 pYTK036
Chloramphenicol
D / 1 pYTK037
Chloramphenicol
D / 2 pYTK038
Chloramphenicol
D / 3 pYTK039
Chloramphenicol
D / 4 pYTK040
Chloramphenicol
D / 5 pYTK041
Chloramphenicol
D / 6 pYTK042
Chloramphenicol
D / 7 pYTK043
Chloramphenicol
D / 8 pYTK044
Chloramphenicol
D / 9 pYTK045
Chloramphenicol
D / 10 pYTK046
Chloramphenicol
D / 11 pYTK047
Chloramphenicol
D / 12 pYTK048
Chloramphenicol
E / 1 pYTK049
Chloramphenicol
E / 2 pYTK050
Chloramphenicol
E / 3 pYTK051
Chloramphenicol
E / 4 pYTK052
Chloramphenicol
E / 5 pYTK053
Chloramphenicol
E / 6 pYTK054
Chloramphenicol
E / 7 pYTK055
Chloramphenicol
E / 8 pYTK056
Chloramphenicol
E / 9 pYTK057
Chloramphenicol
E / 10 pYTK058
Chloramphenicol
E / 11 pYTK059
Chloramphenicol
E / 12 pYTK060
Chloramphenicol
F / 1 pYTK061
Chloramphenicol
F / 2 pYTK062
Chloramphenicol
F / 3 pYTK063
Chloramphenicol
F / 4 pYTK064
Chloramphenicol
F / 5 pYTK065
Chloramphenicol
F / 6 pYTK066
Chloramphenicol
F / 7 pYTK067
Chloramphenicol
F / 8 pYTK068
Chloramphenicol
F / 9 pYTK069
Chloramphenicol
F / 10 pYTK070
Chloramphenicol
F / 11 pYTK071
Chloramphenicol
F / 12 pYTK072
Chloramphenicol
G / 1 pYTK073
Chloramphenicol
G / 2 pYTK074
Chloramphenicol
G / 3 pYTK075
Chloramphenicol
G / 4 pYTK076
Chloramphenicol
G / 5 pYTK077
Chloramphenicol
G / 6 pYTK078
Chloramphenicol
G / 7 pYTK079
Chloramphenicol
G / 8 pYTK080
Chloramphenicol
G / 9 pYTK081
Chloramphenicol
G / 10 pYTK082
Chloramphenicol
G / 11 pYTK083
Ampicillin
G / 12 pYTK084
Kanamycin
H / 1 pYTK085
Spectinomycin
H / 2 pYTK086
Chloramphenicol
H / 3 pYTK087
Chloramphenicol
H / 4 pYTK088
Chloramphenicol
H / 5 pYTK089
Ampicillin
H / 6 pYTK090
Kanamycin
H / 7 pYTK091
Spectinomycin
H / 8 pYTK092
Chloramphenicol
H / 9 pYTK093
Chloramphenicol
H / 10 pYTK094
Chloramphenicol
H / 11 pYTK095
Ampicillin
H / 12 pYTK096
Kanamycin
Data calculated @ 2017-08-21

Kit Plate Map - #1000000061

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