Joint Modular Cloning (JMC) Toolkit
(Kit #
1000000233
)
Depositing Lab: Daniel Voytas
The Joint Modular Cloning (JMC) Toolkit includes a modular cloning chassis and set of functional parts for assembling plant transformation vectors. Transformation vectors assembled from these parts can serve numerous experimental applications ranging from overexpression, gene editing, functional genomics, and more complex metabolic engineering and synthetic biology.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
An extensible vector toolkit and parts library for advanced engineering of plant genomes. Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R, Gamo ME, Starker CG, Zhang F, Voytas DF. Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312. PubMed (Link opens in a new window) Article (Link opens in a new window).
Description
The Joint Modular Cloning (JMC) Toolkit is a Golden Gate framework, comprising 75 destination and structural vectors, that supports assembly of arbitrarily complex constructs through a hierarchical and extensible design interface. Key features include full compatibility with the widely-used Phytobrick (MoClo) standard for Level 0 genetic parts; improved capacity and flexibility versus extant kits; and multi-step design paths that rely exclusively on antibiotic counterselections for efficient selection of correct clones. Level 2 destination vectors include domesticated, minimal T-DNA backbones with different origins of replication that support T-DNA development for plant transformation in the context of both binary and ternary Agrobacterium vector systems.
- Figure 1: Hierarchical assembly of constructs from Phytobrick parts. a: New parts are assembled into Level 0 destination vectors using BbsI with the Phytobrick (J1-J11) overhangs. b: Phytobricks can occupy one of 10 atomic part classes, but commonly span simplified classes using J1, J6, J9, and J11. c: Level 0 parts are assembled into Level 1 destination vectors using BsaI with J1 and J11. d: Level 1 modules are assembled into Level 2 destination vectors using PaqCI with a new set of overhangs (M1-MΩ). e: The pMIN vectors are Level 2 destination vectors, including a minimal backbone and 2 binary vector backbones.
- Figure 2: Modular expansion of binary vectors. a: Expansion linkers are Level 1 modules at any position with end-paired Esp3I or BsaI sites. b: Level 2 vectors with expansion linkers can be used directly as transformation constructs. c: Destination donor vector restore a cloning marker and Golden Gate sites into an expansion linker. d: Both BsaI and Esp3I linkers can be used to create Level 2 destination vectors. e: Esp3I linkers can also be used to create Level 1 destination vectors.
Kit Documentation
Additional information may be found at the provided reference, including a full set of supplementary step-by-step cloning protocols.
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 Joint Modular Cloning (JMC) Toolkit was a gift from Daniel Voytas (Addgene kit #1000000233).”
For your Reference section:
An extensible vector toolkit and parts library for advanced engineering of plant genomes. Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R, Gamo ME, Starker CG, Zhang F, Voytas DF. Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312. PubMed (Link opens in a new window) Article (Link opens in a new window).
Joint Modular Cloning (JMC) Toolkit - #1000000233
- 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 and Kanamycin | |
| Chloramphenicol and Ampicillin | |
| Spectinomycin | |
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pMIN-0 | Chloramphenicol and Kanamycin |
| A / 2 | pMIN-VS1 | Chloramphenicol and Kanamycin |
| A / 3 | pMIN-Ri | Chloramphenicol and Kanamycin |
| A / 4 | pJMC-1T | Chloramphenicol and Ampicillin |
| A / 5 | pJMC-1 | Chloramphenicol and Ampicillin |
| A / 6 | pJMC-2T | Chloramphenicol and Ampicillin |
| A / 7 | pJMC-2 | Chloramphenicol and Ampicillin |
| A / 8 | pJMC-3T | Chloramphenicol and Ampicillin |
| A / 9 | pJMC-3 | Chloramphenicol and Ampicillin |
| A / 10 | pJMC-4T | Chloramphenicol and Ampicillin |
| A / 11 | pJMC-4 | Chloramphenicol and Ampicillin |
| A / 12 | pJMC-5T | Chloramphenicol and Ampicillin |
| B / 1 | pJMC-5 | Chloramphenicol and Ampicillin |
| B / 2 | pJMC-6T | Chloramphenicol and Ampicillin |
| B / 3 | pJMC-6 | Chloramphenicol and Ampicillin |
| B / 4 | pJMC-7T | Chloramphenicol and Ampicillin |
| B / 5 | pJMC-1T-Reverse | Chloramphenicol and Ampicillin |
| B / 6 | pJMC-1-Reverse | Chloramphenicol and Ampicillin |
| B / 7 | pJMC-2T-Reverse | Chloramphenicol and Ampicillin |
| B / 8 | pJMC-2-Reverse | Chloramphenicol and Ampicillin |
| B / 9 | pJMC-3T-Reverse | Chloramphenicol and Ampicillin |
| B / 10 | pJMC-3-Reverse | Chloramphenicol and Ampicillin |
| B / 11 | pJMC-4T-Reverse | Chloramphenicol and Ampicillin |
| B / 12 | pJMC-4-Reverse | Chloramphenicol and Ampicillin |
| C / 1 | pJMC-5T-Reverse | Chloramphenicol and Ampicillin |
| C / 2 | pJMC-5-Reverse | Chloramphenicol and Ampicillin |
| C / 3 | pJMC-6T-Reverse | Chloramphenicol and Ampicillin |
| C / 4 | pJMC-6-Reverse | Chloramphenicol and Ampicillin |
| C / 5 | pJMC-7T-Reverse | Chloramphenicol and Ampicillin |
| C / 6 | pJMC-L2-Donor-BsaI | Chloramphenicol and Ampicillin |
| C / 7 | pJMC-L2-Donor-Esp3I | Chloramphenicol and Ampicillin |
| C / 8 | pJMC-L1-Donor-Esp3I | Chloramphenicol and Ampicillin |
| C / 9 | pJMC-L2-Reverse-Donor-BsaI | Chloramphenicol and Ampicillin |
| C / 10 | pJMC-L2-Reverse-Donor-Esp3I | Chloramphenicol and Ampicillin |
| C / 11 | pJMC-L1-Reverse-Donor-Esp3I | Chloramphenicol and Ampicillin |
| C / 12 | pMC-1-ccdB-6 | Spectinomycin |
| D / 1 | pMC-6-ccdB-9 | Spectinomycin |
| D / 2 | pMC-9-ccdB-11 | Spectinomycin |
| D / 3 | pMC-1-ccdB-4 | Spectinomycin |
| D / 4 | pMC-1-ccdB-11 | Spectinomycin |
| D / 5 | pJMC-1-BsaI | Ampicillin |
| D / 6 | pJMC-2T-BsaI | Ampicillin |
| D / 7 | pJMC-2-BsaI | Ampicillin |
| D / 8 | pJMC-3T-BsaI | Ampicillin |
| D / 9 | pJMC-3-BsaI | Ampicillin |
| D / 10 | pJMC-4T-BsaI | Ampicillin |
| D / 11 | pJMC-4-BsaI | Ampicillin |
| D / 12 | pJMC-5T-BsaI | Ampicillin |
| E / 1 | pJMC-5-BsaI | Ampicillin |
| E / 2 | pJMC-6T-BsaI | Ampicillin |
| E / 3 | pJMC-6-BsaI | Ampicillin |
| E / 4 | pJMC-7T-BsaI | Ampicillin |
| E / 5 | pJMC-1-Esp3I | Ampicillin |
| E / 6 | pJMC-2T-Esp3I | Ampicillin |
| E / 7 | pJMC-2-Esp3I | Ampicillin |
| E / 8 | pJMC-3T-Esp3I | Ampicillin |
| E / 9 | pJMC-3-Esp3I | Ampicillin |
| E / 10 | pJMC-4T-Esp3I | Ampicillin |
| E / 11 | pJMC-4-Esp3I | Ampicillin |
| E / 12 | pJMC-5T-Esp3I | Ampicillin |
| F / 1 | pJMC-5-Esp3I | Ampicillin |
| F / 2 | pJMC-6T-Esp3I | Ampicillin |
| F / 3 | pJMC-6-Esp3I | Ampicillin |
| F / 4 | pJMC-7T-Esp3I | Ampicillin |
| F / 5 | pJMC-1-Empty | Ampicillin |
| F / 6 | pJMC-2T-Empty | Ampicillin |
| F / 7 | pJMC-2-Empty | Ampicillin |
| F / 8 | pJMC-3T-Empty | Ampicillin |
| F / 9 | pJMC-3-Empty | Ampicillin |
| F / 10 | pJMC-4T-Empty | Ampicillin |
| F / 11 | pJMC-4-Empty | Ampicillin |
| F / 12 | pJMC-5T-Empty | Ampicillin |
| G / 1 | pJMC-5-Empty | Ampicillin |
| G / 2 | pJMC-6T-Empty | Ampicillin |
| G / 3 | pJMC-6-Empty | Ampicillin |
| G / 4 | pJMC-7T-Empty | Ampicillin |