Voytas Lab - MoClo Plant Parts III: Transformation & Genome Engineering Kit
(Kit #
1000000236
)
Depositing Lab: Daniel Voytas
The MoClo Plant Parts III: Transformation & Genome Engineering Kit contains 94 Level 0 parts, compatible with the Phytobrick/MoClo system for hierarchical golden gate assembly of plant transformation constructs. All parts are domesticated for BsaI and PaqCI, and the majority are also domesticated for Esp3I, BbsI, and SapI sites.
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 kit includes regulatory elements, coding sequences, basic CRISPR parts, and parts for modular geminiviral replicon assembly.
- Regulatory elements: 21 promoters (J1-J4 or J1-J6 part classes), including constitutive, housekeeping, viral, and inducible promoters for both monocot and dicot expression, 19 monocot and dicot terminators (J9-J11), 5 enhancer-blocking insulator sequences (J1-J11), and 1 hairpin transcription insulator sequence (J1-J4)
- Coding sequences (J6-J9): 8 different site-specific recombinases, 5 fluorescent reporters (GFP, RFP, CFP), 4 luciferase reporters (Firefly, Renilla, Emerald, RedF), 2 metabolic reporters (Ruby, fungal luciferase), 2 selectable markers (HptII, NptII), 5 developmental regulators (IPT, AtPLT5, TaGRF-GIF, ZmBbm, ZmWus2), and AtSpCas9
- CRISPR reagents: Arabidopsis and rice U6 promoters and SpCas9 guide scaffold, for direct assembly of sgRNA expression cassettes into Level 1 module
- Geminiviral replicons: parts for Wheat Dwarf Virus (WDV) and Bean Yellow Dwarf Virus (BeYDV), including long intergenic region (LIR), short intergenic region (SIR), and replication initiator protein (Rep)/RepA sequences, compatible with the JMC Toolkit (Addgene Kit # 1000000233) for multigene assembly
Kit Documentation
Protocols and supporting materials: Detailed cloning protocols for use of these parts, as well as expression benchmarking data for many of the regulatory elements, available as part of the associated publication (Chamness et al. 2023).
Related materials: For construct assembly resources, see also the JMC Toolkit and other Phytobrick-compatible cloning toolkits (MoClo, GoldenBraid, Mobius) available on Addgene. For additional Phytobrick parts libraries, see MoClo Plant Parts, MoClo Plant Parts II, MoClo CRISPR/Cas Toolkit, and others.
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 Voytas Lab MoClo Plant Parts III: Transformation & Genome Engineering Kit was a gift from Daniel Voytas (Addgene kit #1000000236).”
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).
MoClo Plant Parts III: Transformation & Genome Engineering Kit - #1000000236
- 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
Spectinomycin |
Inventory
Well | Plasmid | Resistance |
---|---|---|
A / 1 | pMC-1-EXOB-11 | Spectinomycin |
A / 2 | pMC-1-TBS-11 | Spectinomycin |
A / 3 | pMC-1-Ugi1-11 | Spectinomycin |
A / 4 | pMC-1-Ugi3-11 | Spectinomycin |
A / 5 | pMC-1-Ugi4-11 | Spectinomycin |
A / 6 | pMC-1-Hairpin_Insulator-4 | Spectinomycin |
A / 7 | pMC-1-CmYLCV-4 | Spectinomycin |
A / 8 | pMC-1-DR5v2-4 | Spectinomycin |
A / 9 | pMC-1-OsEf1a-4 | Spectinomycin |
A / 10 | pMC-1-Nos-6 | Spectinomycin |
A / 11 | pMC-1-35S_Long-6 | Spectinomycin |
A / 12 | pMC-1-CmYLCV-6 | Spectinomycin |
B / 1 | pMC-1-AtUbi10-6 | Spectinomycin |
B / 2 | pMC-1-GmHSP17-5E-6 | Spectinomycin |
B / 3 | pMC-1-NtEf1a-6 | Spectinomycin |
B / 4 | pMC-1-OsEf1a-6 | Spectinomycin |
B / 5 | pMC-1-OsUbi-6 | Spectinomycin |
B / 6 | pMC-1-PvUbi1-6 | Spectinomycin |
B / 7 | pMC-1-PvUbi2-6 | Spectinomycin |
B / 8 | pMC-1-SlEf1a-6 | Spectinomycin |
B / 9 | pMC-1-SlRPS5A1-6 | Spectinomycin |
B / 10 | pMC-1-SlUbi-6 | Spectinomycin |
B / 11 | pMC-1-StUbi-6 | Spectinomycin |
B / 12 | pMC-1-ZmAXIG1-6 | Spectinomycin |
C / 1 | pMC-1-ZmEf1a-6 | Spectinomycin |
C / 2 | pMC-1-ZmPLTP-6 | Spectinomycin |
C / 3 | pMC-1-ZmUbi1-6 | Spectinomycin |
C / 4 | pMC-6-BeYDV-Rep-RepA-9 | Spectinomycin |
C / 5 | pMC-6-WDV-Rep-RepA-9 | Spectinomycin |
C / 6 | pMC-1-BeYDV-LS-L-11 | Spectinomycin |
C / 7 | pMC-1-BeYDV-LRS-L-11 | Spectinomycin |
C / 8 | pMC-1-WDV-LS-L-11 | Spectinomycin |
C / 9 | pMC-1-WDV-LRS-L-11 | Spectinomycin |
C / 10 | pMC-6-Cre-9 | Spectinomycin |
C / 11 | pMC-6-Cre-StLS1-9 | Spectinomycin |
C / 12 | pMC-6-Cre-GR-9 | Spectinomycin |
D / 1 | pMC-6-FLPe-9 | Spectinomycin |
D / 2 | pMC-6-PhiC31-9 | Spectinomycin |
D / 3 | pMC-6-R-9 | Spectinomycin |
D / 4 | pMC-6-Integrase4-9 | Spectinomycin |
D / 5 | pMC-6-Integrase13-9 | Spectinomycin |
D / 6 | pMC-6-CinH-9 | Spectinomycin |
D / 7 | pMC-6-Bxb1-9 | Spectinomycin |
D / 8 | pMC-6-AmCyan-9 | Spectinomycin |
D / 9 | pMC-6-Cycle3GFP-9 | Spectinomycin |
D / 10 | pMC-6-EGFP-9 | Spectinomycin |
D / 11 | pMC-6-StayGold-9 | Spectinomycin |
D / 12 | pMC-6-DsRed2-9 | Spectinomycin |
E / 1 | pMC-6-ELuc-9 | Spectinomycin |
E / 2 | pMC-6-FLuc-9 | Spectinomycin |
E / 3 | pMC-6-RedFLuc-9 | Spectinomycin |
E / 4 | pMC-6-RLuc-9 | Spectinomycin |
E / 5 | pMC-6-Ruby-9 | Spectinomycin |
E / 6 | pMC-6-GFP-P2A-NptII-9 | Spectinomycin |
E / 7 | pMC-6-AmCyan-P2A-NptII-9 | Spectinomycin |
E / 8 | pMC-6-HptII-9 | Spectinomycin |
E / 9 | pMC-6-NptII-9 | Spectinomycin |
E / 10 | pMC-6-AtPLT5-9 | Spectinomycin |
E / 11 | pMC-6-IPT-9 | Spectinomycin |
E / 12 | pMC-6-TaGRF-GIF-9 | Spectinomycin |
F / 1 | pMC-6-ZmBbm-9 | Spectinomycin |
F / 2 | pMC-6-ZmWus2-9 | Spectinomycin |
F / 3 | pMC-6-ZmBbm-P2A-ZmWus2-9 | Spectinomycin |
F / 4 | pMC-6-AsNPGA-9 | Spectinomycin |
F / 5 | pMC-6-NnCPH-9 | Spectinomycin |
F / 6 | pMC-6-NnH3H-9 | Spectinomycin |
F / 7 | pMC-6-NnHISPS-9 | Spectinomycin |
F / 8 | pMC-6-NnLuz-9 | Spectinomycin |
F / 9 | pMC-6-HpaB-9 | Spectinomycin |
F / 10 | pMC-6-HpaC-9 | Spectinomycin |
F / 11 | pMC-6-RcTAL-9 | Spectinomycin |
F / 12 | pMC-9-35S-11 | Spectinomycin |
G / 1 | pMC-9-AtHSP-11 | Spectinomycin |
G / 2 | pMC-9-NbAct3-11 | Spectinomycin |
G / 3 | pMC-9-NbHSP-11 | Spectinomycin |
G / 4 | pMC-9-NtEU-11 | Spectinomycin |
G / 5 | pMC-9-NtEf1a-11 | Spectinomycin |
G / 6 | pMC-9-OsAct2-11 | Spectinomycin |
G / 7 | pMC-9-OsEf1a-11 | Spectinomycin |
G / 8 | pMC-9-OsT28-11 | Spectinomycin |
G / 9 | pMC-9-OsUbi-11 | Spectinomycin |
G / 10 | pMC-9-PinII-11 | Spectinomycin |
G / 11 | pMC-9-RBCS-11 | Spectinomycin |
G / 12 | pMC-9-SlEf1a-11 | Spectinomycin |
H / 1 | pMC-9-SlRPS5A1-11 | Spectinomycin |
H / 2 | pMC-9-SlUbi-11 | Spectinomycin |
H / 3 | pMC-9-StUbi-11 | Spectinomycin |
H / 4 | pMC-9-ZmEf1a-11 | Spectinomycin |
H / 5 | pMC-9-ZmIn2-11 | Spectinomycin |
H / 6 | pMC-9-ZmUbi-11 | Spectinomycin |
H / 7 | pMC-6-AtSpCas9-9 | Spectinomycin |
H / 8 | pMC-1-AtU6-Y | Spectinomycin |
H / 9 | pMC-1-OsU6-X | Spectinomycin |
H / 10 | pMC-Z-SpCas9-gRNA-repeat-polyT-11 | Spectinomycin |