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 93 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.
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 93 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 standard basic parts (including promoters, terminators, operators, transcriptional activators/repressors and protein coding regions) and 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.
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
Plasmid Sequences
Sequences for kit plasmids (CSV format)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 PMID 26839579 .
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 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
| Chloramphenicol | |
| Ampicillin | |
| Kanamycin | |
| Spectinomycin |