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.

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.

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: YTK summary 128.0 KB

Download a ZIP file containing all MoClo-YTK plasmid sequences in GenBank format: 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

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 @ 2024-04-19