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Orzaez Lab GoldenBraid 2.0 Kit
(Kit # 1000000076 )

Depositing Lab:   Diego Orzaez

The GoldenBraid 2.0 Kit provides plant scientists with a resource for the modular cloning of multigenic DNA constructs in Plant Synthetic Biology applications. The kit consists of 95 plasmids which include the complete set of vectors required to perform GoldenBraid cloning. Detailed information, tutorials and software tools for GB cloning can be found at https://gbcloning.upv.es.

All Golden Braid 2.0 DNA parts are fully compatible with Golden Braid 3.0.

This kit will be sent as bacterial glycerol stocks in 96-well plate format.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Sarrion-Perdigones A, Vazquez-Vilar M, Palací J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D. Plant Physiol. 2013 Jul;162(3):1618-31. PubMed PMID 23669743 .

Description

The GoldenBraid 2.0 Kit is a collection of 95 plasmids which includes the complete set of vectors required to perform GoldenBraid cloning:

  • pUPD2 - a universal domesticator plasmid for cloning of basic DNA parts
  • pDGB1 (based on the pGreenII backbone) and pDGB3 (based on pCambia1302 backbone) - two series of GoldenBraid destination plasmids for the construction of transcriptional units/modules for plant expression
  • four twister plasmids to swap transcriptional units or modules between destination plasmids

It also includes 51 standard basic parts (including promoters, terminators, operators, transcriptional activators/repressors and protein coding regions) and 17 transcriptional units with different functions as resistance markers, reporter genes, CRISPR/Cas9 genome editing tools, etc.

In addition, the kit incorporates a set of parts to allow for the construction of CRISPR/Cas9 constructs expressing multiple gRNAs (in a monocistronic or polycistronic strategy) to be used for gene editing or transcriptional regulation. Detailed information, tutorials and software tools for GB cloning can be found at https://gbcloning.upv.es

All Golden Braid 2.0 DNA parts are fully compatible with Golden Braid 3.0.

Orzaez Lab flowchart of Golden Braid Kit

Depositor-provided Files

The Orzaez lab has generated a webpage providing support for GoldenBraid users, including searchable databases of GB elements and experiments and a number of software tools for in silico simulation of DNA assembly reactions. GB cloning webpage

Additional References

Software-Assisted Stacking of Gene Modules Using GoldenBraid 2.0 DNA-Assembly Framework. Vazquez-Vilar M, Sarrion-Perdigones A, Ziarsolo P, Blanca J, Granell A, Orzaez D. Methods Mol Biol 2015; 1284:399-420. PubMed .

Design and construction of multigenic constructs for plant biotechnology using the GoldenBraid cloning strategy. Sarrion-Perdigones A, Palaci J, Granell A, Orzaez D. Methods Mol Biol 2014; 1116:133-51. PubMed .

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 GoldenBraid 2.0 kit was a gift from Diego Orzaez (Addgene kit # 1000000076 ).”

For your Reference section:

GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Sarrion-Perdigones A, Vazquez-Vilar M, Palací J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D. Plant Physiol. 2013 Jul;162(3):1618-31. PubMed PMID 23669743 .

A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Vazquez-Vilar M, Bernabé-Orts JM, Fernandez-Del-Carmen A, Ziarsolo P, Blanca J, Granell A, Orzaez D. Plant Methods 2016 Feb 1;12:10. PubMed .

GoldenBraid 2.0 Kit - #1000000076

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 links to its kit plate map on the right. 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
Kanamycin
Spectinomycin

Inventory

Well Plasmid Resistance
A / 1 pUPD2
Chloramphenicol
A / 2 pPAt2S3 (GB0029)
Ampicillin
A / 3 pP35S (GB0030)
Ampicillin
A / 4 pPML1 (GB0045)
Ampicillin
A / 5 pNH (GB0064)
Ampicillin
A / 6 pPnos (GB0072)
Ampicillin
A / 7 pPMtb (GB0080)
Ampicillin
A / 8 pP2A11 (GB0082)
Ampicillin
A / 9 pPTctp (GB0083)
Ampicillin
A / 10 pPUbc (GB0145)
Ampicillin
A / 11 pHsp18.2 (GB0153)
Ampicillin
A / 12 pHsp70B (GB0155)
Ampicillin
B / 1 p2x35S (GB0222)
Ampicillin
B / 2 pPAtUbq10 (GB0223)
Ampicillin
B / 3 pPromE8 (GB0914)
Ampicillin
B / 4 pUPD_Mini35s(-60,UTR) (GB0050)
Ampicillin
B / 5 pPAct2noUTR (GB0192)
Ampicillin
B / 6 p5'UTRAct2 (GB0193)
Ampicillin
B / 7 pOpLacI:mini35ss (GB0534)
Ampicillin
B / 8 pOpLexA::mini35s (GB0733)
Ampicillin
B / 9 pLexAOp (GB0118)
Ampicillin
B / 10 p6xUAS (GB0179)
Ampicillin
B / 11 pLacIBD::SDRX (GB0673)
Ampicillin
B / 12 pLexA::SRDX (GB0732)
Ampicillin
C / 1 pGal4BD::BDR (GB0734)
Ampicillin
C / 2 pTHsp18.2 (GB0035)
Ampicillin
C / 3 pT35S (GB0036)
Ampicillin
C / 4 pTnos (GB0037)
Ampicillin
C / 5 pT2A11 (GB0143)
Ampicillin
C / 6 pTE8 (GB0144)
Ampicillin
C / 7 pTAct2 (GB0210)
Ampicillin
C / 8 pTAtUbq3 (GB0219)
Ampicillin
C / 9 pBFP (GB0025)
Ampicillin
C / 10 pRosea (GB0026)
Ampicillin
C / 11 pP19 (GB0038)
Ampicillin
C / 12 pHcPro (GB0067)
Ampicillin
D / 1 pYFP (GB0053)
Ampicillin
D / 2 pRenilla (GB0056)
Ampicillin
D / 3 pMyb12 (GB0075)
Ampicillin
D / 4 pDelila (GB0079)
Ampicillin
D / 5 pLuciferase (GB0096)
Ampicillin
D / 6 pDsRed (GB0100)
Ampicillin
D / 7 pGUS (GB0208)
Ampicillin
D / 8 pHygromycin (GB0211)
Ampicillin
D / 9 phumanCas9 (GB0575)
Ampicillin
D / 10 pD10AH840AhCas9 (GB1041)
Ampicillin
D / 11 pU6-26 (GB1001)
Ampicillin
D / 12 pUPD_OsU3 (GB1184)
Ampicillin
E / 1 psgRNA (GB0645)
Ampicillin
E / 2 pTNos:BASTA:PNos (GB0023)
Ampicillin
E / 3 pEGB Tnos:NptII:Pnos (GB0226)
Kanamycin
E / 4 pEGB 1alfa1R Tnos:Hygromycin:PNos (GB0235)
Kanamycin
E / 5 pDGB3alpha2_35S:P19:Tnos (GB1203)
Kanamycin
E / 6 pEGB 35S:Renilla:Tnos (GB0109)
Kanamycin
E / 7 pEGB 35S:Luciferase:Tnos (GB0110)
Kanamycin
E / 8 pEGB Pnos:Luciferase:Tnos (GB0112)
Kanamycin
E / 9 pEGB3 Pnos:Luciferase:Tnos-SF-35S:Renilla:Tnos-35S:P19:Tnos-SF (GB1116)
Kanamycin
E / 10 pEGB 35S:YFP:Tnos (GB0209)
Kanamycin
E / 11 pEGB 35S:DsRed:Tnos (GB0361)
Kanamycin
E / 12 pEGB2alpha2 35s:hCas9:tNos (GB0639)
Kanamycin
F / 1 pEGB 35S:dCas9:Tnos (GB1191)
Kanamycin
F / 2 pDGB1_alpha1
Kanamycin
F / 3 pDGB1_alpha2
Kanamycin
F / 4 pDGB1_alpha1R
Kanamycin
F / 5 pDGB1_alpha2R
Kanamycin
F / 6 pDGB3_alpha1
Kanamycin
F / 7 pDGB3_alpha2
Kanamycin
F / 8 pDGB3_alpha1R
Kanamycin
F / 9 pDGB3_alpha2R
Kanamycin
F / 10 pDGB1alpha1_SF (GB0106)
Kanamycin
F / 11 pDGB1alpha2_SF (GB0107)
Kanamycin
F / 12 pDGB1_omega1
Spectinomycin
G / 1 pDGB1_omega2
Spectinomycin
G / 2 pDGB1_omega1R
Spectinomycin
G / 3 pDGB1_omega2R
Spectinomycin
G / 4 pDGB3_omega1
Spectinomycin
G / 5 pDGB3_omega2
Spectinomycin
G / 6 pDGB3_omega1R
Spectinomycin
G / 7 pDGB3_omega2R
Spectinomycin
G / 8 pDGB1omega1_SF (GB0158)
Spectinomycin
G / 9 pDGB1omega2_SF (GB0466)
Spectinomycin
G / 10 pdCas9 (GB1079)
Ampicillin
G / 11 pEGB2Ω2_SF-35s:hCas9:tNos (GB1103)
Spectinomycin
G / 12 pEGB 35s:dCas:BRD:tNos (GB1172)
Kanamycin
H / 1 pEGB 35s:dCas:VP64:tNos (GB1189)
Kanamycin
H / 2 pEGB 35s:dCas:EDLL:tNos (GB1190)
Kanamycin
H / 3 pU6-1 (GB1204)
Chloramphenicol
H / 4 tRNA-gRNA position [D1_2] (GB1205)
Ampicillin
H / 5 tRNA-gRNA position [2_n-1] (GB1206)
Ampicillin
H / 6 tRNA-gRNA position [n] (GB1207)
Ampicillin
H / 7 tRNA-gRNA position [D1_n-1] (GB1208)
Ampicillin
H / 8 tRNA-gRNA position [M1_2] (GB1209)
Ampicillin
H / 9 tRNA-gRNA position [M1_n-1] (GB1210)
Ampicillin
H / 10 pEGB SF-35S:Renilla:TNOS-35S:P19:TNOS (GB0160)
Kanamycin
H / 11 pEGB3omega1R Tnos:NptII:Pnos-SF (GB1181)
Spectinomycin
Data calculated @ 2017-05-22

Kit Plate Map - #1000000076

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