Yeast GoldenBraid Cloning System and Toolkit
(Kit #
1000000138
)
Depositing Lab: Elena Caro Bernat
The Yeast GoldenBraid Cloning System and Toolkit provides modular pieces (promoters, MTSs, 8xHis N terminal tag, terminators and resistance cassettes) for the generation of transcriptional units using the GoldenBraid cloning system. This toolkit allows for integration in the S. cerevisiae genome at two different loci supporting high and stable transgene expression: YPRCΔ15 and YORWΔ22.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria Pérez-González A., Kniewel R., Veldhuizen M., Verma H.K., Navarro-Rodríguez M., Rubio L.M., & Caro E. BMC Biotechnology. (2017).17(1), 80; doi: 10.1186/s12896-017-0393-y. PubMed PMID: 29132331.
Description
The Yeast GoldenBraid Cloning System and Toolkit contains:
- 4 GoldenBraid adapted vectors for the cloning of transcriptional units and their integration in the yeast genome (S. cerevisiae W303 or CEN.PK) at locus YPRCΔ15
- 4 adapted vectors for integration into the yeast genome YORWΔ22 locus
- 9 vectors containing 8 constitutive and 1 inducible promoter
- pUPD-pGAL1 (01-11)
- pUPD-pTDH3 (01-11)
- pUPD-pTDH2 (01-11)
- pUPD-pTEF2 (01-11)
- pUPD-pHXT7 (01-11)
- pUPD-pTPI1 (01-11)
- pUPD-pPYK1 (01-11)
- pUPD-PGK1 (01-11)
- pUPD-PGI1 (01-11)
- 8 vectors containing mitochondrial targeting signals (MTSs)
- pUPD-Su9MTS (12)
- pUPD-SOD2MTS (12)
- pUPD-MAM33MTS (12)
- pUPD2-ODPAMTS (12)
- pUPD2-ODPBMTS (12)
- pUPD2-ATPAMTS (12)
- pUPD2-GLRX2MTS (12)
- pUPD-MTS2 (12)
- 1 vector for 8xHis N terminal tagging pUC57-8xHis
- Vectors containing 3 terminators and 2 resistance cassettes (Hygromycin resistance and G418 resistance)
Schematic representation of the adaptation of GoldenBraid vectors for yeast chromosome integration.
5’- and 3’- AOH represents YPRCΔ15 or YORWΔ22. Numbers 1, 2 and 3 indicate the overhangs generated after digestion. AOH: arms of homology.
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 Yeast GoldenBraid Cloning System and Toolkit was a gift from Elena Caro Bernat (Addgene kit #1000000138).”
For your Reference section:
Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria Pérez-González A., Kniewel R., Veldhuizen M., Verma H.K., Navarro-Rodríguez M., Rubio L.M., & Caro E. BMC Biotechnology. (2017).17(1), 80; doi: 10.1186/s12896-017-0393-y. PubMed PMID: 29132331.
Yeast GoldenBraid Cloning System and Toolkit - #1000000138
- 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 | |
| Chloramphenicol | |
| Spectinomycin | |
| Kanamycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pUPD-pGAL1 (01-11) | Ampicillin |
| A / 2 | pUPD-pTDH3 (01-11) | Ampicillin |
| A / 3 | pUPD-pTDH2 (01-11) | Ampicillin |
| A / 4 | pUPD-pTEF2 (01-11) | Ampicillin |
| A / 5 | pUPD-pHXT7 (01-11) | Ampicillin |
| A / 6 | pUPD-pTPI1 (01-11) | Ampicillin |
| A / 7 | pUPD-pPYK1 (01-11) | Ampicillin |
| A / 8 | pUPD-PGK1 (01-11) | Ampicillin |
| A / 9 | pUPD-PGI1 (01-11) | Ampicillin |
| A / 10 | pUPD-Su9MTS (12) | Ampicillin |
| A / 11 | pUPD-SOD2MTS (12) | Ampicillin |
| A / 12 | pUPD-MAM33MTS (12) | Ampicillin |
| B / 1 | pUPD2-ODPAMTS (12) | Chloramphenicol |
| B / 2 | pUPD2-ODPBMTS (12) | Chloramphenicol |
| B / 3 | pUPD2-ATPAMTS (12) | Chloramphenicol |
| B / 4 | pUPD2-GLRX2MTS (12) | Chloramphenicol |
| B / 5 | pUPD-MTS2 (12) | Ampicillin |
| B / 6 | pUC57-8xHis | Ampicillin |
| B / 7 | pUPD-Cyc1t (17-21) | Ampicillin |
| B / 8 | pUPD-ADH2t (17-21) | Ampicillin |
| B / 9 | pUPD-TDH2t (17-21) | Ampicillin |
| B / 10 | pUPD-G418R (01-21) | Ampicillin |
| B / 11 | pUPD-HygroBR (01-21) | Ampicillin |
| B / 12 | YORWΔ22Ω2 | Spectinomycin |
| C / 1 | YORWΔ22Ω1 | Spectinomycin |
| C / 2 | YORWΔ22α2 | Kanamycin |
| C / 3 | YORWΔ22α1 | Kanamycin |
| C / 4 | YPRCΔ15Ω2 | Spectinomycin |
| C / 5 | YPRCΔ15Ω1 | Spectinomycin |
| C / 6 | YPRCΔ15α2 | Kanamycin |
| C / 7 | YPRCΔ15α1 | Kanamycin |