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Addgene

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.

Add to Cart
$400 USD + shipping
Available to academics and nonprofits only.

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)
Data calculated @ 2024-10-31

Kit Plate Map - #1000000133

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