GeneArt

1 | | Life Technologies Proprietary Confidential | 3/17/2011 Life Technologies Proprietary & & Confidential |


Evolution of the Genomics Market
Before the Genome
Hypothesis driven Single organism
One geneone protein

Sequenced Genomes
Systems biology
Pathway & networks Functional correlation

Synthetic Design
Deterministic design
Optimized living systems Metabolic engineering

Discover
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Develop

Produce

Healthcare

Biofuels

Chemicals

Agriculture

BioRemediation

Enabling new therapies for cancer and other diseases

Harnessing photosynthet ic organisms to produce fuels directly from CO2

Replacing chemical processes by green, biobased processes

Developing high-yield, more resistant and economic plant feedstocks

Developing microbial solutions to clean water and generate energy in the process

3 | Life Technologies Proprietary & Confidential | 3/17/2011 Genome Synthesis and Design Futures Implications for the US Economy DOE Bio-Era report - Feb 2007

Life Technologies

DNA Parts & Devices

BioDesign Software

Generating synthetic solutions to improve the human condition

Specialty Media

Optimized Hosts

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GENEART:
GENEART , 2011Life Technologies
GENEART Life Technologies GENEART 1.

2. DNA

Synthetic Biology leader in gene synthesis State-of-the-art gene design & optimization Scale & capacity for rapid delivery

Leader in cloning, DNA assembly & vector design Skill & reach in sequencing Commercial excellence

Leading tool providers for Synthetic Biology applications

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GENEART
DNA ISO

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Bioinforma tics

Oligo Synthesis

Gene/ome Production

Sequencing & QC

Laboratory Information Management System

Security / Safety High throughput, minidisc-nanoscale, oligo-libraries automated assembly, directed evolution, cGMP products 100% fidelity

Optimization / Design

Frontload production
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DNA
Save money -- Predictable cloning experiments means 0% rework
Save time -- Research productivity gain; PCR cloning should not be trial and error

Convenience -- 100% sequence fidelity by ordering online

Better performance -- Overcome DNA design and expression limitations via optimized genes

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GeneOptimizer
Up to x100 fold improvement Wild Type / Natural Gene
GeneOptimizer

Expression of HIV-1 gag in HEK293 cells.

Optimized Synthetic Gene

GeneOptimizer patent JP 4510640 granted in Japan 2010
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in various expression systems

Reference projects
US National Cancer Institute (NCI): Within the scope of the project, GENEART produced more than 3,700 genes for the completion of the Mammalian Gene Collection. HuCAL Platinum: New HuCAL PLATINUM antibody library is based on DNA components synthesized by GENEART Welcome Trust Sanger Institute: The Wellcome Trust Sanger Institute awards a major contract on synthesizing up to 600 genes to GENEART, used for identification of mechanisms involved in genetic and infectious diseases.

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GENEART

DNA cDNA UTRs

1.
2. 3.

4.

4DNA 10DNA Web-based in silico assembly and DNA oligo design tool

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GENEART Seamless Cloning and Assembly Kit

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15 bp Terminal Homology Requirement

Fragment 1 Fragment 2

Terminal homology enables simultaneous and directional cloning of PCR fragments. Homology can be obtained in multiple ways.
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Ordering Information

Product
GENEART Seamless Cloning and Assembly Kit GENEART Linear pUC19L Vector for Seamless Cloning Web-based DNA Oligo Designer (invitrogen.com/DesignDNAassembly)

Quantity
20 rxns 20 rxns

Cat. No.
A13288 A13289

For more information and to place an order, please visit: www.invitrogen.com/DNAassembly

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GENEART High-Order Genetic Assembly System

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Terminal Homology Requirement

Fragment 1 Fragment 2

Fragments end-homology

Oligo Stitching (with synthetic DNA linkers)

40

40

Terminal homology enables simultaneous and directional cloning of up to 10 DNA fragments. Homology can be obtained in multiple ways, including oligo stitching.
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Oligo Stitching DNA

Perfect Junctions

Ideal for elements that are pre-existing or too big to PCR

Imperfect Junctions

Ideal for sequence editing

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Highly Efficient Cloning of Large Fragments

GENEART High Order Genetic Assembly System has a high cloning efficiency even as the number of fragments cloned approaches 10 (total size of 100 kbp).
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Ordering Information
Product GENEART High-Order Genetic Assembly System GENEART High-Order Genetic Assembly Systems (with Yeast Growth Media) GENEART High-Order Linear pYES1L Vector CSM Media for Mav203 Yeast Cells GENEART High-Order Vector Conversion Cassette Web-based DNA Oligo Designer (invitrogen.com/DesignDNAassembly) Quantity 10 rxns 10 rxns 2L media 10 rxns 2 liter 10 rxns Cat. No. A13285 A13286 A13287 A13292 A13291 -

For more information and to place an order, please visit: www.invitrogen.com/DNAassembly

19 | Life Technologies Proprietary & Confidential | 3/17/2011

DNA
GENEART Seamless Cloning & Assembly GENEART High Order Genetic Assembly

In vitro

In vivo

Up to 4 fragments plus vector

15 bp overlaps Constructs up to 13 kbp

Up to 10 fragments plus vector

30 bp overlaps Constructs up to 110 kbp

Seamless (no scars)

On-line design tools

Seamless (no scars)

On-line design tools

Single tube 30 min reaction

Bridging oligo option: reuse and edit

Yeast- E. coli transfer (10 min), no liquid culture

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DNA
Designs DNA oligos for PCR and/or oligo stitching Checks for potential issues during assembly Delivers final construction in GenBank format

www.invitrogen.com/DesignDNAassembly

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GENEART Site-Directed Mutagenesis System

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GENEART
, , and method to generate mutant constructs in vitro

Substitutions, deletions, or insertions of up to 12 High efficiency of mutagenesis;

nucleotides in plasmids up to 14 Kb

over 90% for a 3 Kb plasmid less than 3 hours to complete

Time savings though a protocol that takes

Flexible system that works for plasmids of many sizes, with DNA isolated from any
source and with no need for specialized vectors, host strains, or restriction sites
Product Quantity 16 rxns Cat. No. A13282

GENEART Site-Directed Mutagenesis System

For more information and to place an order, please visit: www.invitrogen.com/DNAassembly

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Mutagenesis Workflow

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Mutagenesis Workflow (Cont.)

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2010 Life Technologies. All Rights Reserved. Proprietary 2010 Life Technologies. All Rights Reserved. Proprietary and Confidential. and Co

Mutagenesis Efficiency vs. Competitor

GENEART System has a higher mutagenesis efficiency as the size of the construct increases
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For More Information

www.invitrogen.com/DNAassembly

Genetic Engineering & Biotechnology News

Vol 30, Number 18 Oct. 15, 2010

Check out our live and on-demand webinars available at www.invitrogen.com/DNAassembly

27 | Life Technologies Proprietary & Confidential | 3/17/2011