Voytas Lab - Golden Gate TALEN and TAL Effector Kit 2.0
(Kit #
1000000024
)
Depositing Labs: Daniel Voytas, Adam Bogdanove
This kit and accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays, containing anywhere between 12 and 31 repeats.
This kit consists of one 96-well plate, and will be shipped as bacterial glycerol stocks on dry ice. Samples should be frozen at -80°C immediately upon arrival. Individual plasmids can be ordered from each plasmid page and will be shipped as bacterial stabs.
Original Publication
Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. PubMed 21493687
Description
TALENs are fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease. TALENs bind and cleave DNA in pairs, and their binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. This collection and accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays, containing anywhere between 12 and 31 repeats.
In January 2013, additional plasmids were added to the Golden Gate kit. They include the 11 NH RVD module plasmids. Streubel et al. (2012) and Cong et al. (2012) show that the RVD NH targets G more specifically than NN does, with good apparent affinity. NK also targets G specifically, but with lower affinity (Christian et al., 2012). NH is therefore the currently preferred RVD for targeting G.
pZHY500 and pZHY501 are Golden Gate compatible yeast expression vectors with deletions in the coding sequence that result in truncations of both the amino and carboxyl portions of the TAL portion of the protein to match the Miller et al., 2011 architecture. Details can be found in Plant Physiol. 2013 Jan;161(1):20-7.
pCP5b is an unpublished plasmid that allows one to test TALEN activity in yeast. This plasmid is very similar to and can be used in place of the published pCP5 (see Proc Natl Acad Sci U S A. 107(26):12028-33). You can find a detailed protocol on how to design and clone targets for the Yeast TALEN assay: Yeast TALEN assay 52.9Â KB
Kit Documentation
GG 2.0 kit.pdf 104.2Â KBResources
Web based software to search for TAL binding sites
Protocols
Golden Gate TALEN assembly protocol 645.4Â KBHow 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 plasmid kit used for generation of TALENs was a gift from Daniel Voytas and Adam Bogdanove (Addgene kit # 1000000024)"
For your Reference section:
Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Apr 14. PubMed 21493687
Golden Gate TALEN and TAL effector kit 2.0 - #1000000024
- 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
| Tetracycline | |
| Spectinomycin | |
| Ampicillin | |
| Chloramphenicol and Ampicillin | |
| Chloramphenicol |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pHD1 | Tetracycline |
| A / 2 | pHD2 | Tetracycline |
| A / 3 | pHD3 | Tetracycline |
| A / 4 | pHD4 | Tetracycline |
| A / 5 | pHD5 | Tetracycline |
| A / 6 | pHD6 | Tetracycline |
| A / 7 | pHD7 | Tetracycline |
| A / 8 | pHD8 | Tetracycline |
| A / 9 | pHD9 | Tetracycline |
| A / 10 | pHD10 | Tetracycline |
| A / 11 | pNG1 | Tetracycline |
| A / 12 | pNG2 | Tetracycline |
| B / 1 | pNG3 | Tetracycline |
| B / 2 | pNG4 | Tetracycline |
| B / 3 | pNG5 | Tetracycline |
| B / 4 | pNG6 | Tetracycline |
| B / 5 | pNG7 | Tetracycline |
| B / 6 | pNG8 | Tetracycline |
| B / 7 | pNG9 | Tetracycline |
| B / 8 | pNG10 | Tetracycline |
| B / 9 | pNI1 | Tetracycline |
| B / 10 | pNI2 | Tetracycline |
| B / 11 | pNI3 | Tetracycline |
| B / 12 | pNI4 | Tetracycline |
| C / 1 | pNI5 | Tetracycline |
| C / 2 | pNI6 | Tetracycline |
| C / 3 | pNI7 | Tetracycline |
| C / 4 | pNI8 | Tetracycline |
| C / 5 | pNI9 | Tetracycline |
| C / 6 | pNI10 | Tetracycline |
| C / 7 | pNN1 | Tetracycline |
| C / 8 | pNN2 | Tetracycline |
| C / 9 | pNN3 | Tetracycline |
| C / 10 | pNN4 | Tetracycline |
| C / 11 | pNN5 | Tetracycline |
| C / 12 | pNN6 | Tetracycline |
| D / 1 | pNN7 | Tetracycline |
| D / 2 | pNN8 | Tetracycline |
| D / 3 | pNN9 | Tetracycline |
| D / 4 | pNN10 | Tetracycline |
| D / 5 | pNK1 | Tetracycline |
| D / 6 | pNK2 | Tetracycline |
| D / 7 | pNK3 | Tetracycline |
| D / 8 | pNK4 | Tetracycline |
| D / 9 | pNK5 | Tetracycline |
| D / 10 | pNK6 | Tetracycline |
| D / 11 | pNK7 | Tetracycline |
| D / 12 | pNK8 | Tetracycline |
| E / 1 | pNK9 | Tetracycline |
| E / 2 | pNK10 | Tetracycline |
| E / 3 | pNH1 | Tetracycline |
| E / 4 | pNH2 | Tetracycline |
| E / 5 | pNH3 | Tetracycline |
| E / 6 | pNH4 | Tetracycline |
| E / 7 | pNH5 | Tetracycline |
| E / 8 | pNH6 | Tetracycline |
| E / 9 | pNH7 | Tetracycline |
| E / 10 | pNH8 | Tetracycline |
| E / 11 | pNH9 | Tetracycline |
| E / 12 | pNH10 | Tetracycline |
| F / 1 | pLR-HD | Spectinomycin |
| F / 2 | pLR-NG | Spectinomycin |
| F / 3 | pLR-NI | Spectinomycin |
| F / 4 | pLR-NN | Spectinomycin |
| F / 5 | pFUS_B1 | Spectinomycin |
| F / 6 | pFUS_B2 | Spectinomycin |
| F / 7 | pFUS_B3 | Spectinomycin |
| F / 8 | pFUS_B4 | Spectinomycin |
| F / 9 | pFUS_B5 | Spectinomycin |
| F / 10 | pFUS_B6 | Spectinomycin |
| F / 11 | pFUS_B7 | Spectinomycin |
| F / 12 | pFUS_B8 | Spectinomycin |
| G / 1 | pFUS_B9 | Spectinomycin |
| G / 2 | pFUS_B10 | Spectinomycin |
| G / 3 | pFUS_A | Spectinomycin |
| G / 4 | pFUS_A30A | Spectinomycin |
| G / 5 | pFUS_A30B | Spectinomycin |
| G / 6 | pLR-NH | Spectinomycin |
| G / 7 | pTAL1 | Ampicillin |
| G / 8 | pTAL2 | Ampicillin |
| G / 9 | pTAL3 | Ampicillin |
| G / 10 | pTAL4 | Ampicillin |
| G / 11 | pZHY500 | Ampicillin |
| G / 12 | pZHY501 | Ampicillin |
| H / 1 | pCP5b | Chloramphenicol and Ampicillin |
| H / 2 | pKEB31 | Chloramphenicol |