GoldenPiCS Kit
(Kit #
1000000133
)
Depositing Lab: Brigitte Gasser, Diethard Mattanovich, Michael Sauer
GoldenPiCS is a plasmid toolkit which provides a flexible modular system for advanced strain engineering in P. pastoris (based on the multi-organism system GoldenMOCS). This kit allows users flexible generation of overexpression plasmids with up to eight transcription units. The kit contains 19 different promoters and 10 transcription terminators, four selection markers and four loci for targeted genome integration.
This kit will be sent as bacterial glycerol stocks in a 96-well plate format.
Original Publication
GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris. Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, Sauer M, Mattanovich D, Gasser B, Marx H. BMC Sys Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3. PubMed PMID 29221460 .
Description
GoldenPiCS is a Golden Gate-derived Pichia pastoris cloning system (based on the multi-organism system GoldenMOCS- see image below). The GoldenPiCS Kit was deposited by the Mattanovich/Gasser/Sauer Group from BOKU Vienna University and contains three hierarchical backbone (BB) levels for flexible generation of overexpression plasmids with up to eight transcription units. The kit contains 19 different promoters and 10 transcription terminators, four selection markers and four loci for targeted genome integration. Both promoters and terminators are in BB1 plasmids with different fusion sites. These single parts (promoter, CDS, terminator) are then assembled into a recipient BB2 plasmid to form a functional expression cassette. BB3 plasmids serve to assemble multiple expression cassettes (e.g. a whole pathway) into one vector (see image below).
This plasmid toolkit can be used for strain engineering of P. pastoris to accomplish pathway expression, protein production or other applications where the integration of various DNA products is required.
Image from Prielhofer et al., BMC Sys Biol, 2017 Dec 8;11(1):123.
Plasmid Sequences
Sequences for kit plasmids (CSV format)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 GoldenPiCS Kit was a gift from the Gasser/Mattanovich/Sauer Group (Addgene kit #1000000133)"
For your Reference section:
GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris. Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, Sauer M, Mattanovich D, Gasser B, Marx H. BMC Sys Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3. PubMed PMID 29221460 .
GoldenPiCS Kit - #1000000133
- 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
| Kanamycin | |
| Ampicillin | |
| Bleocin (Zeocin) | |
| Hygromycin | |
| Nourseothricin (clonNat) |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | BB1_12 | Kanamycin |
| A / 2 | BB1_23 | Kanamycin |
| A / 3 | BB1_34 | Kanamycin |
| A / 4 | BB1_12_pGAP | Kanamycin |
| A / 5 | BB1_12_pGPM1 | Kanamycin |
| A / 6 | BB1_12_pRPP1B | Kanamycin |
| A / 7 | BB1_12_pPDC1 | Kanamycin |
| A / 8 | BB1_12_pPOR1 | Kanamycin |
| A / 9 | BB1_12_pMDH3 | Kanamycin |
| A / 10 | BB1_12_pADH2 | Kanamycin |
| A / 11 | BB1_12_pFBA1-1 | Kanamycin |
| A / 12 | BB1_12_pSHB17 | Kanamycin |
| B / 1 | BB1_12_pTEF2 | Kanamycin |
| B / 2 | BB1_12_pRPL2A | Kanamycin |
| B / 3 | BB1_12_pLAT1 | Kanamycin |
| B / 4 | BB1_12_pPFK300 | Kanamycin |
| B / 5 | BB1_12_pGUT1 | Kanamycin |
| B / 6 | BB1_12_pTHI11 | Kanamycin |
| B / 7 | BB1_12_pDAS2 | Kanamycin |
| B / 8 | BB1_12_pAOX1 | Kanamycin |
| B / 9 | BB1_12_pDAS1 | Kanamycin |
| B / 10 | BB1_12_pFDH1 | Kanamycin |
| B / 12 | BB1_23_eGFP | Kanamycin |
| C / 1 | BB1_34_ScCYC1tt | Kanamycin |
| C / 2 | BB1_34_TDH3tt | Kanamycin |
| C / 3 | BB1_34_RPS2tt | Kanamycin |
| C / 4 | BB1_34_RPP1Btt | Kanamycin |
| C / 5 | BB1_34_RPS17Btt | Kanamycin |
| C / 6 | BB1_34_chr4_0883tt | Kanamycin |
| C / 7 | BB1_34_IDP1tt | Kanamycin |
| C / 8 | BB1_34_RPS25Att | Kanamycin |
| C / 9 | BB1_34_RPS3tt | Kanamycin |
| C / 10 | BB1_34_RPL2Att | Kanamycin |
| C / 11 | BB3aK_14* | Kanamycin |
| C / 12 | BB3aK_AC | Kanamycin |
| D / 1 | BB3aK_AD | Kanamycin |
| D / 2 | BB3aK_AE | Kanamycin |
| D / 3 | BB3aK_AF | Kanamycin |
| D / 4 | BB2_AB | Ampicillin |
| D / 5 | BB2_BC | Ampicillin |
| D / 6 | BB2_CD | Ampicillin |
| D / 7 | BB2_DE | Ampicillin |
| D / 8 | BB2_EF | Ampicillin |
| D / 9 | BB2_FG | Ampicillin |
| D / 10 | BB2_GH | Ampicillin |
| D / 11 | BB2_HI | Ampicillin |
| D / 12 | BB3aZ_14* | Bleocin (Zeocin) |
| E / 1 | BB3eH_14* | Hygromycin |
| E / 2 | BB3eH_AC | Hygromycin |
| E / 3 | BB3eH_AD | Hygromycin |
| E / 4 | BB3eH_AE | Hygromycin |
| E / 5 | BB3eH_AF | Hygromycin |
| E / 6 | BB3aN_14* | Nourseothricin (clonNat) |
| E / 7 | BB3eN_14* | Nourseothricin (clonNat) |
| E / 8 | BB3rN_14* | Nourseothricin (clonNat) |
| E / 9 | BB3rN_AC | Nourseothricin (clonNat) |
| E / 10 | BB3rN_AD | Nourseothricin (clonNat) |
| E / 11 | BB3rN_AE | Nourseothricin (clonNat) |
| E / 12 | BB3rN_AF | Nourseothricin (clonNat) |
| F / 1 | BB3rN_AG | Nourseothricin (clonNat) |
| F / 2 | BB3rN_AH | Nourseothricin (clonNat) |
| F / 3 | BB3rN_AI | Nourseothricin (clonNat) |