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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 at $65 per plasmid, and will be shipped as bacterial stabs.

Add to Cart
$425 USD + shipping

Available to academics and nonprofits only.

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

Description

TALEN s 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pCP5b is an unpublished plasmid that allows one to test TALEN activity in yeast.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: Icon Yeast TALEN assay (52.9 KB)

Kit Documentation

Icon GG 2.0 kit.pdf (104.2 KB)

Resources

Web based software to search for TAL binding sites

Icon TALEN sequencing guide (76.3 KB)

Icon Plasmid sequences and maps (4.8 MB)

Protocols

Icon Golden Gate TALEN assembly protocol (645.4 KB)

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 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

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 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

Tetracycline
Spectinomycin
Ampicillin (50 ug/ml)
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 (50 ug/ml)
G / 8 pTAL2
Ampicillin (50 ug/ml)
G / 9 pTAL3
Ampicillin (50 ug/ml)
G / 10 pTAL4
Ampicillin (50 ug/ml)
G / 11 pZHY500
Ampicillin
G / 12 pZHY501
Ampicillin
H / 1 pCP5b
Chloramphenicol and Ampicillin
H / 2 pKEB31
Chloramphenicol
Data calculated @ 2016-12-05

Kit Plate Map - #1000000024

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