MoClo Yeast Secretion and Display (YSD) Toolkit
(Kit #
1000000240
)
Depositing Lab: Paul Young
The Yeast Secretion and Display Toolkit is a collection of 34 plasmids that can be used in conjunction with the MoClo Yeast Toolkit (YTK) to optimize secretion and cell surface display of proteins of interest. It enables the efficient incorporation of the coding sequences for a panel of seventeen well-characterized secretion-promoting sequences and five surface display anchor proteins into Saccharomyces cerevisiae expression cassettes.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization of Heterologous Protein Secretion and Surface Display in Saccharomyces cerevisiae. O'Riordan NM, Juric V, O'Neill SK, Roche AP, Young PW. ACS Synth Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743. PubMed (Link opens in a new window) Article (Link opens in a new window).
Description
The MoClo YSD toolkit provides an efficient system to rapidly sample small, carefully curated libraries of secretion-promoting sequences and/or surface display anchors for a recombinant protein of interest. The kit contains plasmids encoding the following S. cerevisiae secretion-promoting parts: nine pre-signal peptide (SP) sequences, four mating factor-α pre-pro-SP sequences (wild-type and three engineered derivatives) and four translational fusion partner (TFP) sequences. In addition, it includes parts encoding four yeast (Aga2p, Cwp2p, Sed1p and Tip1p) and one synthetic (649 stalk) surface display anchors. These parts can be combined with promoters, terminators, epitope and affinity tags, as well as selectable markers and replication origins from the MoClo Yeast Toolkit (YTK) and Multiplex Yeast Toolkit (MYT) developed by the Dueber and Ellis labs respectively. Used in this way, the MoClo YSD toolkit harnesses the efficiencies of the modular cloning framework in order to standardize the trial-and-error process of expression cassette generation, thereby accelerating the design, build, test cycle for synthetic biology experiments involving protein secretion or surface display in yeast.
The kit also contains plasmids encoding two proteins of interest for use as controls. These are the fluorescent proteins, mRuby2 and S. cerevisiae invertase (SUC2). Coding sequences for these two proteins are included as type 3b' parts and as expression plasmids (without a SP - for intracellular expression, and with an Aga2 pre-SP or SSCW4 TFP – for secretion). An expression plasmid for cell surface display of mRuby2 with the SED1 anchor is also included.
- Figure 1. Schematic overview of the MoClo Yeast Secretion and Display (YSD) Toolkit. (A) Summary of the secretion-promoting sequences (SPS) and surface display anchor parts in the MoClo YSD toolkit (blue and purple respectively). (B) MoClo YSD parts are compatible with the MoClo Yeast Toolkit (YTK) and Multiplex Yeast Toolkit (MYT), which provide the part types indicated. (C) The panels of SPS and anchor proteins in the YSD Toolkit can be readily incorporated into yeast expression constructs by modular cloning to rapidly identify the optimal sequences for secretion and/or surface display of a protein of interest (POI). HA = HA-epitope tag, SP = Signal Peptide.
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 Yeast Secretion and Display (YSD) Toolkit was a gift from Paul Young (Addgene kit #1000000240).”
For your Reference section:
A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization of Heterologous Protein Secretion and Surface Display in Saccharomyces cerevisiae. O'Riordan NM, Juric V, O'Neill SK, Roche AP, Young PW. ACS Synth Biol. ACS Synth Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743. PubMed (Link opens in a new window) Article (Link opens in a new window).
MoClo Yeast Secretion and Display (YSD) Toolkit - #1000000240
- 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
| Chloramphenicol | |
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pYSD001 | Chloramphenicol |
| A / 2 | pYSD002 | Chloramphenicol |
| A / 3 | pYSD003 | Chloramphenicol |
| A / 4 | pYSD004 | Chloramphenicol |
| A / 5 | pYSD005 | Chloramphenicol |
| A / 6 | pYSD006 | Chloramphenicol |
| A / 7 | pYSD007 | Chloramphenicol |
| A / 8 | pYSD008 | Chloramphenicol |
| A / 9 | pYSD009 | Chloramphenicol |
| A / 10 | pYSD021 | Chloramphenicol |
| A / 11 | pYSD022 | Chloramphenicol |
| A / 12 | pYSD023 | Chloramphenicol |
| B / 1 | pYSD024 | Chloramphenicol |
| B / 2 | pYSD041 | Chloramphenicol |
| B / 3 | pYSD042 | Chloramphenicol |
| B / 4 | pYSD043 | Chloramphenicol |
| B / 5 | pYSD044 | Chloramphenicol |
| B / 6 | pYSD061 | Chloramphenicol |
| B / 7 | pYSD063 | Chloramphenicol |
| B / 8 | pYSD080 | Chloramphenicol |
| B / 9 | pYSD081 | Chloramphenicol |
| B / 10 | pYSD082 | Chloramphenicol |
| B / 11 | pYSD083 | Chloramphenicol |
| B / 12 | pYSD084 | Chloramphenicol |
| C / 1 | pYSD085 | Chloramphenicol |
| C / 2 | pYSD099 | Ampicillin |
| C / 3 | pYSD100 | Ampicillin |
| C / 4 | pYSD101 | Ampicillin |
| C / 5 | pYSD113 | Ampicillin |
| C / 6 | pYSD140 | Ampicillin |
| C / 7 | pYSD141 | Ampicillin |
| C / 8 | pYSD153 | Ampicillin |
| C / 9 | pYSD300 | Ampicillin |
| C / 10 | pYSD313 | Ampicillin |