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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 -80oC immediately upon arrival. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs.

$425 USD + shipping
Available to Academic 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

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pCP5b is an unpublished plasmid that allows one to test TALEN activity in yeast.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: 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

Resistance Color Key:

 
  Tetracycline  
 
  Spectinomycin  
 
  Ampicillin (50 ug/ml)  
 
  Ampicillin  
 
  Ampicillin and Chloramphenicol  
 
  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
  Ampicillin and Chloramphenicol
H / 2 pKEB31
  Chloramphenicol
Data from: CALCULATED @ 2014-11-27 06:56:24.979066