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Zinc Finger Consortium: Modular Assembly Kit
(Kit # 1000000005 )

Depositing Labs:   Keith Joung, Dan Voytas

Zinc finger nucleases (ZFNs) are synthetic proteins commonly used in genome engineering that contain an custom-designed zinc finger DNA-binding domain fused to the cleavage domain of the FokI endonuclease. This kit includes 141 zinc finger plasmids encoding different zinc finger modules from the Sangamo, Barbas and Toolgen module sets, and can be used to efficiently assemble zinc finger arrays via modular assembly. Also included in this kit also are 5 plasmids and a strain for the bacterial two-hybrid (B2H) system used to screen zinc finger arrays for activity. To request only the B2H reagents, please see the Zinc Finger Consortium: Modular Assembly Accessory Reagents Kit (#1000000010).

This kit consists of 2 plates, and will be shipped as bacterial glycerol stocks, 96-well format, 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
$650 USD + shipping

Available to academics and nonprofits only.

Original Publication

Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc . Nat Protoc. 2006;1(3):1637-52. PubMed .

Description

Zinc fingers are the most common family of transcription factors in organisms ranging from yeast to humans. Custom zinc finger proteins can be designed to target a specific DNA sequence in a gene of interest in nearly any cell type.

Zinc finger nucleases (ZFNs), which consist of an engineered zinc finger DNA-binding domain fused to the DNA cleavage domain of the FokI enzyme, are powerful tools for performing genome engineering in a variety of organisms. ZFNs can introduce double-stranded breaks in DNA that stimulate both homologous and non-homologous recombination pathways to alter a desired locus in the genome.

The zinc finger modules in this kit were derived from the Barbas, Sangamo and ToolGen module sets and standardized to permit "mixing-and-matching" of the different modules. A complementary web-based Zinc Finger Targeter program (ZiFiT) was designed to assist in the identification of potential DNA target sites.

In May 2008, the laboratories of Keith Joung, Daniel Voytas, and Toni Cathomen published a large-scale evaluation of the modular assembly method and reported important guidance for researchers interested in using this approach to engineer multi-finger domains. The work is described in:

Unexpected failure rates for modular assembly of engineered zinc fingers. Ramirez CL, Foley JE, Wright DA, Müller-Lerch F, Rahman SH, Cornu TI, Winfrey RJ, Sander JD, Fu F, Townsend JA, Cathomen T, Voytas DF, Joung JK. Nat Methods . 2008 May;5(5):374-5. doi: 10.1038/nmeth0508-374

Kit Documentation

Icon ZFC Expression Vector and Modular Assembly Kit Info (113.7 KB)

Links

In July 2012, a legacy version of the ZiFiT software package (ZiFiT v3.3) was made available to maintain support for modular assembly of zinc fingers using the Modular Assembly Kit developed by the Zinc Finger Consortium (Wright et al., Nature Protocols 1, 1637-1652, 2006). Considerable progress has been made towards engineering custom DNA binding proteins and the Zinc Finger Consortium now strongly recommends utilizing newer alternative methods for engineering DNA binding domains. For more information on these methods please see the current version of ZiFiT .

The legacy version, ZiFiT (v3.3) , can be used to identify and engineer zinc finger proteins.

Resources

A spreadsheet containing information for the 141 zinc finger plasmids is available as a CSV file:

Icon Zinc Finger Plasmid sequences (761.6 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 engineering ZFNs was a gift from Keith Joung and Dan Voytas (Addgene kit # 1000000005)"

For your Reference section:
Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK. Nat Protoc . Nat Protoc. 2006;1(3):1637-52. PubMed .

Zinc Finger Consortium Expression Vector and Modular Assembly Kit - #1000000005

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

Ampicillin
Kanamycin
Chloramphenicol
Tetracycline

Inventory

Well Plasmid Resistance
Plate 1 / A / 1 pc3XB-ZF1
Ampicillin
Plate 1 / A / 2 pc3XB-ZF2
Ampicillin
Plate 1 / A / 3 pc3XB-ZF3
Ampicillin
Plate 1 / A / 4 pc3XB-ZF4
Ampicillin
Plate 1 / A / 5 pc3XB-ZF5
Ampicillin
Plate 1 / A / 6 pc3XB-ZF6
Ampicillin
Plate 1 / A / 7 pc3XB-ZF7
Ampicillin
Plate 1 / A / 8 pc3XB-ZF8
Ampicillin
Plate 1 / A / 9 pc3XB-ZF9
Ampicillin
Plate 1 / A / 10 pc3XB-ZF10
Ampicillin
Plate 1 / A / 11 pc3XB-ZF11
Ampicillin
Plate 1 / A / 12 pc3XB-ZF12
Ampicillin
Plate 1 / B / 1 pc3XB-ZF13
Ampicillin
Plate 1 / B / 2 pc3XB-ZF14
Ampicillin
Plate 1 / B / 3 pc3XB-ZF15
Ampicillin
Plate 1 / B / 4 pc3XB-ZF16
Ampicillin
Plate 1 / B / 5 pc3XB-ZF17
Ampicillin
Plate 1 / B / 6 pc3XB-ZF18
Ampicillin
Plate 1 / B / 7 pc3XB-ZF19
Ampicillin
Plate 1 / B / 8 pc3XB-ZF20
Ampicillin
Plate 1 / B / 9 pc3XB-ZF21
Ampicillin
Plate 1 / B / 10 pc3XB-ZF22
Ampicillin
Plate 1 / B / 11 pc3XB-ZF23
Ampicillin
Plate 1 / B / 12 pc3XB-ZF24
Ampicillin
Plate 1 / C / 1 pc3XB-ZF25
Ampicillin
Plate 1 / C / 2 pc3XB-ZF26
Ampicillin
Plate 1 / C / 3 pc3XB-ZF27
Ampicillin
Plate 1 / C / 4 pc3XB-ZF28
Ampicillin
Plate 1 / C / 5 pc3XB-ZF29
Ampicillin
Plate 1 / C / 6 pc3XB-ZF30
Ampicillin
Plate 1 / C / 7 pc3XB-ZF31
Ampicillin
Plate 1 / C / 8 pc3XB-ZF32
Ampicillin
Plate 1 / C / 9 pc3XB-ZF33
Ampicillin
Plate 1 / C / 10 pc3XB-ZF34
Ampicillin
Plate 1 / C / 11 pc3XB-ZF35
Ampicillin
Plate 1 / C / 12 pc3XB-ZF36
Ampicillin
Plate 1 / D / 1 pc3XB-ZF37
Ampicillin
Plate 1 / D / 2 pc3XB-ZF38
Ampicillin
Plate 1 / D / 3 pc3XB-ZF39
Ampicillin
Plate 1 / D / 4 pc3XB-ZF40
Ampicillin
Plate 1 / D / 5 pc3XB-ZF41
Ampicillin
Plate 1 / D / 6 pc3XB-ZF42
Ampicillin
Plate 1 / D / 7 pc3XB-ZF43
Ampicillin
Plate 1 / D / 8 pc3XB-ZF44
Ampicillin
Plate 1 / D / 9 pc3XB-ZF45
Ampicillin
Plate 1 / D / 10 pc3XB-ZF46
Ampicillin
Plate 1 / D / 11 pc3XB-ZF47
Ampicillin
Plate 1 / D / 12 pc3XB-ZF48
Ampicillin
Plate 1 / E / 1 pc3XB-ZF49
Ampicillin
Plate 1 / E / 2 pc3XB-ZF50
Ampicillin
Plate 1 / E / 3 pc3XB-ZF51
Ampicillin
Plate 1 / E / 4 pc3XB-ZF52
Ampicillin
Plate 1 / E / 5 pc3XB-ZF53
Ampicillin
Plate 1 / E / 6 pc3XB-ZF54
Ampicillin
Plate 1 / E / 7 pc3XB-ZF55
Ampicillin
Plate 1 / E / 8 pc3XB-ZF56
Ampicillin
Plate 1 / E / 9 pc3XB-ZF57
Ampicillin
Plate 1 / E / 10 pc3XB-ZF58
Ampicillin
Plate 1 / E / 11 pc3XB-ZF59
Ampicillin
Plate 1 / E / 12 pc3XB-ZF60
Ampicillin
Plate 1 / F / 1 pc3XB-ZF61
Ampicillin
Plate 1 / F / 2 pc3XB-ZF62
Ampicillin
Plate 1 / F / 3 pc3XB-ZF63
Ampicillin
Plate 1 / F / 4 pc3XB-ZF64
Ampicillin
Plate 1 / F / 5 pc3XB-ZF65
Ampicillin
Plate 1 / F / 6 pc3XB-ZF66
Ampicillin
Plate 1 / F / 7 pc3XB-ZF67
Ampicillin
Plate 1 / F / 8 pc3XB-ZF68
Ampicillin
Plate 1 / F / 9 pc3XB-ZF69
Ampicillin
Plate 1 / F / 10 pc3XB-ZF70
Ampicillin
Plate 1 / F / 11 pc3XB-ZF71
Ampicillin
Plate 1 / F / 12 pc3XB-ZF72
Ampicillin
Plate 1 / G / 1 pc3XB-ZF73
Ampicillin
Plate 1 / G / 2 pc3XB-ZF74
Ampicillin
Plate 1 / G / 3 pc3XB-ZF75
Ampicillin
Plate 1 / G / 4 pc3XB-ZF76
Ampicillin
Plate 1 / G / 5 pc3XB-ZF77
Ampicillin
Plate 1 / G / 6 pc3XB-ZF78
Ampicillin
Plate 1 / G / 7 pc3XB-ZF79
Ampicillin
Plate 1 / G / 8 pc3XB-ZF80
Ampicillin
Plate 1 / G / 9 pc3XB-ZF81
Ampicillin
Plate 1 / G / 10 pc3XB-ZF82
Ampicillin
Plate 1 / G / 11 pc3XB-ZF83
Ampicillin
Plate 1 / G / 12 pc3XB-ZF84
Ampicillin
Plate 1 / H / 1 pc3XB-ZF85
Ampicillin
Plate 1 / H / 2 pc3XB-ZF86
Ampicillin
Plate 1 / H / 3 pc3XB-ZF87
Ampicillin
Plate 1 / H / 4 pc3XB-ZF88
Ampicillin
Plate 1 / H / 5 pc3XB-ZF89
Ampicillin
Plate 1 / H / 6 pc3XB-ZF90
Ampicillin
Plate 1 / H / 7 pc3XB-ZF91
Ampicillin
Plate 1 / H / 8 pc3XB-ZF92
Ampicillin
Plate 1 / H / 9 pc3XB-ZF93
Ampicillin
Plate 1 / H / 10 pc3XB-ZF94
Ampicillin
Plate 1 / H / 11 pc3XB-ZF95
Ampicillin
Plate 1 / H / 12 pc3XB-ZF96
Ampicillin
Plate 2 / A / 1 pc3XB-ZF97
Ampicillin
Plate 2 / A / 2 pc3XB-ZF98
Ampicillin
Plate 2 / A / 3 pc3XB-ZF99
Ampicillin
Plate 2 / A / 4 pc3XB-ZF100
Ampicillin
Plate 2 / A / 5 pc3XB-ZF101
Ampicillin
Plate 2 / A / 6 pc3XB-ZF102
Ampicillin
Plate 2 / A / 7 pc3XB-ZF103
Ampicillin
Plate 2 / A / 8 pc3XB-ZF104
Ampicillin
Plate 2 / A / 9 pc3XB-ZF105
Ampicillin
Plate 2 / A / 10 pc3XB-ZF106
Ampicillin
Plate 2 / A / 11 pc3XB-ZF107
Ampicillin
Plate 2 / A / 12 pc3XB-ZF108
Ampicillin
Plate 2 / B / 1 pc3XB-ZF109
Ampicillin
Plate 2 / B / 2 pc3XB-ZF110
Ampicillin
Plate 2 / B / 3 pc3XB-ZF111
Ampicillin
Plate 2 / B / 4 pc3XB-ZF112
Ampicillin
Plate 2 / B / 5 pc3XB-ZF113
Ampicillin
Plate 2 / B / 6 pc3XB-ZF114
Ampicillin
Plate 2 / B / 7 pc3XB-ZF115
Ampicillin
Plate 2 / B / 8 pc3XB-ZF116
Ampicillin
Plate 2 / B / 9 pc3XB-ZF117
Ampicillin
Plate 2 / B / 10 pc3XB-ZF118
Ampicillin
Plate 2 / B / 11 pc3XB-ZF119
Ampicillin
Plate 2 / B / 12 pc3XB-ZF120
Ampicillin
Plate 2 / C / 1 pc3XB-ZF121
Ampicillin
Plate 2 / C / 2 pc3XB-ZF122
Ampicillin
Plate 2 / C / 3 pc3XB-ZF123
Ampicillin
Plate 2 / C / 4 pc3XB-ZF124
Ampicillin
Plate 2 / C / 5 pc3XB-ZF125
Ampicillin
Plate 2 / C / 6 pc3XB-ZF126
Ampicillin
Plate 2 / C / 7 pc3XB-ZF127
Ampicillin
Plate 2 / C / 8 pc3XB-ZF128
Ampicillin
Plate 2 / C / 9 pc3XB-ZF129
Ampicillin
Plate 2 / C / 10 pc3XB-ZF130
Ampicillin
Plate 2 / C / 11 pc3XB-ZF131
Ampicillin
Plate 2 / C / 12 pc3XB-ZF132
Ampicillin
Plate 2 / D / 1 pc3XB-ZF133
Ampicillin
Plate 2 / D / 2 pc3XB-ZF134
Ampicillin
Plate 2 / D / 3 pc3XB-ZF135
Ampicillin
Plate 2 / D / 4 pc3XB-ZF136
Ampicillin
Plate 2 / D / 5 pc3XB-ZF137
Ampicillin
Plate 2 / D / 6 pc3XB-ZF138
Ampicillin
Plate 2 / D / 7 pc3XB-ZF139
Ampicillin
Plate 2 / D / 8 pc3XB-ZF140
Ampicillin
Plate 2 / D / 9 pc3XB-ZF141
Ampicillin
Plate 2 / F / 1 pGP-FF
Ampicillin
Plate 2 / F / 2 pGP-FB-orig BA
Ampicillin
Plate 2 / F / 3 pAC-Kan-alphaGal4
Kanamycin
Plate 2 / F / 4 pBAC-lacZ
Chloramphenicol
Plate 2 / F / 5 pBAC-BA-lacZ
Chloramphenicol
Plate 2 / F / 6 KJBAC1 strain
Tetracycline
Plate 2 / F / 7 pST1374
Ampicillin
Plate 2 / F / 8 pDW1775
Ampicillin
Data calculated @ 2017-03-26

Kit Plate Map - #1000000005

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