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Yeast MoClo secretion and surface display toolkit 2.0: improvements and applications for analysis of protein-protein interactions and whole-cell biocatalysis

Jurić V, Erwin LG, O'Riordan NM, Maher E, Holmes JD, Young PW
bioRxiv 2025.08.19.671047 (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
252961pYSD045Encodes S. cerevisiae Pir anchor-linker-HA Tag as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252962pYSD046Encodes S. cerevisiae Pir anchor-linker-2xFLAG Tag as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252963pYSD047Encodes S. cerevisiae Pir2 anchor-linker-MYC Tag as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252964pYSD051Encodes S. cerevisiae Aga2-(NoTag)-linker as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252965pYSD052Encodes S. cerevisiae HA Tag-Aga2-linker as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252966pYSD053Encodes S. cerevisiae 2xFLAG-Tag-Aga2-linker as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252967pYSD054Encodes S. cerevisiae MYC Tag-Aga2-linker as a Type 3a' part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252968pYSD055Encodes S. cerevisiae Aga2-(NoTag)-linker as a Type 3a part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252969pYSD056Encodes S. cerevisiae HA Tag-Aga2-linker as a Type 3a part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252970pYSD057Encodes S. cerevisiae 2xFLAG-Tag-Aga2-linker as a Type 3a part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252971pYSD058Encodes S. cerevisiae MYC Tag-Aga2-linker as a Type 3a part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit to create fusions of proteins of interest to the C-terminus of Aga2
252973pYSD069Encodes Anti-GFP Nanobody (no tag) as a Type 3b' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252974pYSD070Encodes Protein A ZZ domain as a Type 3b' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252978pYSD088Encodes 2xFLAG-Tagged S. cerevisiae Aga2p yeast surface display anchor protein as a Type 4a part to be used in the yeast secretion/display (YSD2.0) MoClo toolkit for N-terminal fusion
252981pYSD091Encodes S. cerevisiae HA-tagged Aga1 del N181aa disp anchor as a Type 4a part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252982pYSD092Encodes S. cerevisiae 2xFLAG-tagged Aga1 del N181aa disp anchor as a Type 4a part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252983pYSD093Encodes MYC-tagged S. cerevisiae Aga1 del N181aa disp anchor as a Type 4a part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252984pYSD094Encodes GFP11-M3-His6-tag as a Type 4a part to be used for fluoresence complementation based detection in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252985pYSD095Encodes GFP11-M3 as a Type 4a part to be used for fluoresence complementation based detection in the yeast secretion/display (YSD2.0) MoClo toolkit Fluoresence Complementation
252988pYSD503Encodes S. cerevisiae ERV29 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252989pYSD504Encodes S. cerevisiae ERJ5 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252990pYSD505Encodes S. cerevisiae ERO1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252991pYSD506Encodes S. cerevisiae HAC1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252992pYSD507Encodes S. cerevisiae MSN4 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252993pYSD508Encodes S. cerevisiae PDI1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252994pYSD509Encodes S. cerevisiae SEB1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252995pYSD510Encodes S. cerevisiae SIL1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252996pYSD511Encodes S. cerevisiae SSO1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252997pYSD512Encodes S. cerevisiae SSO2 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252998pYSD513Encodes S. cerevisiae ERV2 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
252999pYSD514Encodes S. cerevisiae Sec65 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253000pYSD515Encodes S. cerevisiae SRP14 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253001pYSD516Encodes S. cerevisiae SWA2 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253002pYSD517Encodes S. cerevisiae COG5 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253004pYSD519Encodes S. cerevisiae SRP54 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253005pYSD520Encodes S. cerevisiae MNS1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253006pYSD521Encodes S. cerevisiae SEC4 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253007pYSD522Encodes S. cerevisiae CWH41 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253008pYSD523Encodes S. cerevisiae SEC16 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253009pYSD524Encodes S. cerevisiae CYS4 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253010pYSD525Encodes S. cerevisiae SED5 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253011pYSD526Encodes S. cerevisiae JEM1 as a Type 3 part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253012pYSD703Encodes Ideonella sakaiensis MHETase as a Type 3b' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253014pYSD861Encodes Anti-ALFA-tag Nanobody as a Type 3b' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253015pYSD862Encodes Anti-mCherryLaM-4 Nanobody as a Type 3b' part to be used in the yeast secretion/display (YSD2.0) extension of the MoClo yeast toolkit (YTK)
253019pYSD893A MoClo YTK Type 3 part GFP dropout vector to assemble a signal peptide and protein of interest as N-terminal fusion with 2xFLAG-SED1 display anchor in a URA3, CEN6/ARS4 yeast expression plasmid
253021pYSD274Encodes the Aga2 signal peptide, mRuby2 fluorescent protein, C-terminal GFP11-M3 and 6xHistidine tags in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253022pYSD281Encodes the Suc2 signal peptide, Invertase, C-terminal GFP11-M3 and 6xHistidine tags in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253023pYSD288Encodes the S. cerevisiae Aga1 protein (Part3) with pGAL1 inducible promoter, tENO2 terminator, KanamycinR yeast selectable marker and CEN/ARS yeast origin
253024pYSD291Encodes the S. cerevisiae Aga1 protein (Part3) with pPGK1 promoter, tENO2 terminator, KanamycinR yeast selectable marker and CEN/ARS yeast origin
253025pYSD292Encodes the S. cerevisiae Aga1 protein (Part3) with pPGK1 promoter, tENO2 terminator, HygromycinR yeast selectable marker and CEN/ARS yeast origin
253026pYSD293Encodes the S. cerevisiae Aga1 protein (Part3) with ppPGK1 promoter, tENO2 terminator, ZeocinR yeast selectable marker and CEN/ARS yeast origin
253028pYSD295Encodes the S. cerevisiae Aga1 protein (Part3) with pRPL18B promoter, tENO2 terminator, HIS3 selectable marker and CEN/ARS yeast origin
253030pYSD299Encodes the S. cerevisiae Aga1 protein (Part3) with pPGK1 promoter, tENO2 terminator, LEU2 selectable marker and CEN/ARS yeast origin
253031pYSD301Encodes the Aga2 signal peptide, anti GFP nanobody, HA-tagged Aga2p yeast surface display anchor protein with pTEF1 promoter, tTDH1 terminator, LEU2 selectable marker and CEN/ARS yeast origin
253032pYSD310Encodes the MFalpha1 PrePro-app8 signal peptide, Anti-GFP nanobody, HA-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253033pYSD311Encodes the Aga2 signal peptide, mRuby2 fluorescent protein, HA-tagged Aga2p yeast surface display anchor protein with pTEF1 promoter, LEU2 selectable marker
253034pYSD315Encodes the Aga2 signal peptide, mRuby2 fluorescent protein, HA-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253035pYSD316Encodes the MFalpha1 PrePro-app8 signal peptide, Anti-GFP nanobody, HA-tagged Aga1 deltaN181AA yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253037pYSD322Encodes the MFalpha1 PrePro-app8 signal peptide, Anti-HA Frankenbody, MYC-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253038pYSD325Encodes the MFalpha1 PrePro-app8 signal peptide, Protein A ZZ domain, HA-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253039pYSD326Encodes the OST1-pre / MFalpha1 Pro signal peptide, Protein A ZZ domain, HA-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253041pYSD329Encodes the Aga2 signal peptide, anti-mCherry_LaM4 nanobody, HA-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253042pYSD332Encodes HA-tagged Aga2 yeast surface display anchor protein, Anti-GFP nanobody as a C-terminal fusion, in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253043pYSD335Encodes HA-tagged Aga2 with mRuby2 as a C-terminal fusion in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker.
253045pYSD353Encodes the MFalpha1 PrePro-app8 signal peptide, Anti-GFP nanobody, FLAG-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253046pYSD354Encodes the MFalpha1 PrePro-app8 signal peptide, Anti-GFP nanobody, MYC-tagged 649-stalk yeast surface display anchor protein in a yeast expression vector with pTEF1, LEU2 selectable marker
253047pYSD371Encodes an AGA2 Signal Peptide, Anti- ALFAtag_nanobody and 2xFLAG-tagged-649-stalk anchor in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker
253048pYSD373Encodes an AGA2 Signal Peptide, Anti-mCherry_nanobody_LaM-4 and 2xFLAG-tagged-649-stalk anchor in a yeast expression vector with pTEF1 promoter, LEU2 selectable marker

Antibodies from Article