The GeneArt® Gene Synthesis service consists of chemical synthesis, cloning, and sequence verification of virtually any desired genetic sequence. You will receive a bacterial stab and/or purified plasmid containing your synthesized gene—ready for downstream applications. Whether you have limited cloning experience or simply want to save time, the GeneArt® Gene Synthesis service helps you move your ideas from the planning stage to the laboratory more quickly. Benefit from our experience in successfully producing over 180,000 constructs for customers as diverse as large pharmaceutical companies, biotechnology start-ups, and basic research institutions. The comparison shown in the figure below highlights the time and effort saved compared to traditional cloning. For more information visit: https://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cloning/gene-synthesis.html?CID=genesynthesis-SS-12312

1. gene SYnTHeSIS SeRVICeS
headline essence
build your next
headline brow
breakthrough
GeneArt® Gene Synthesis Services

2. geneArt® services—your partner for
gene synthesis through protein production
Whether you need industry-leading gene synthesis services or optimized protein expression, or want to outsource the entire
process from gene synthesis to protein production, this brochure outlines geneArt® services to help you succeed.
Gene synthesis
An increasingly cost-effective method for obtaining DNA constructs with
100% sequence accuracy, GeneArt® services offer:
• Largest capacity and fastest production processes
• Outstanding quality—ISO 9001:2008 certification
• Gene optimization for maximum protein expression
Custom services
Whether you’re looking to save time or improve upon existing processes,
GeneArt® services provide:
• A single resource for all of your outsourcing needs
• Gene synthesis to custom cell line and protein production
• ISO 9001:2008 certification and responsive project management
Contents
Introduction Cell line, protein, and plasmid services
• GeneArt® services overview ......................................... 2 • Cell lines and proteins ................................................. 8
• Plasmid services .......................................................... 9
Gene synthesis and optimization
• Introduction to gene optimization ............................... 3
• Gene synthesis ............................................................. 6
• Directed evolution ........................................................ 7
2 GeneArt® Gene Synthesis Services

3. GeneArt® services by Life Technologies
Considered one of the foremost global specialists in the area of synthetic biology, the geneArt® facility is the world’s leading
manufacturer of synthetic genes.
History
In 1999, Dr. Ralf Wagner, Dr. Marcus Graf, and Dr. Hans Wolf founded
the GeneArt® company after leaving the University of Regensburg.
Dr. Wagner needed custom-built genes to develop vaccines. They were
not commercially available at the time and thus had to be synthesized
de novo. The synthetic genes performed exceptionally well, and the
scientists recognized their potential benefit for research and industry.
They launched GeneArt® services with the vision to synthesize artificial
genes on a large scale at affordable rates, thus enabling access to these
new tools for scientists worldwide.
The GeneArt® company was awarded one of the largest gene synthesis
contracts for completion of the “Mammalian Gene Collection Program”
by the US National Institutes of Health. In addition, they produced
subgenomic elements for the construction of the first synthetic bacterial
genome by the J. Craig Venter Institute.
Customers
The world‘s largest pharmaceutical companies, many international
biotechnology companies, and major universities/research institutes
benefit from using the GeneArt® technology platform. Customers rely on
the expertise and experience of GeneArt® scientists to improve enzymes,
construct genetically altered bacteria, and develop and produce new
therapeutics and vaccines, in addition to providing the highest-quality
synthetic genes.
To learn more and place your order, go to www.invitrogen.com/genesynthesis 3

4. Gene optimization to maximize protein expression
Production of recombinant human proteins in human cells for biomedical research and product development can be
hampered by low expression yields. These expression issues can limit researchers’ ability to conduct structural and
functional analyses, delaying and in some cases halting the discovery process. gene optimization is the solution to
traditional protein expression limitations. The common pain points associated with protein expression—yield, solubility, and
functionality—can now be addressed in a rational and systematic way.
Using data available from published literature in combination with proprietary data, the geneOptimizer® algorithm
determines the optimal gene sequence for your expression experiments (Figure 1). Optimization has been experimentally
proven to increase protein expression rates up to 100-fold in a variety of host systems.
Multigene study of optimized mammalian genes Summary
In a first-of-its-kind study1, five important, biologically relevant protein Following optimization, the 50 genes all showed reliable expression and
classes were selected for study—protein kinases, transcription factors, 86% exhibited elevated expression (Figure 2, page 4). Further analysis
ribosomal proteins, cytokines, and membrane proteins. Then, 50 human showed no detrimental effect on protein solubility and unaltered func-
genes were chosen from the NCBI database to represent the five protein tionality was demonstrated for JNK1, JNK3, and CDC2 using optimized
classes. constructs (data not shown).
The selected genes were individually optimized using the sliding window Using the geneOptimizer® algorithm:
GeneOptimizer® algorithm.2 The corresponding wild type genes were • 96% of optimized genes showed equal or higher protein expression
subcloned using native sequences available from the NCBI database. • Protein yields increased up to 15-fold with optimized genes
Each gene was then prepped and expressed in triplicate in HEK293T • 100% of optimized genes expressed versus 88% of wild type genes
cells.
Sequence repeats Codon usage GC content
PABP
PABP
PABP
AAAAAA AAAAAA
Killer motifs Splice sites RNA secondary structures
Optimized gene
Figure 1. Proprietary GeneOptimizer ® algorithm determines optimal gene sequence for your experiments.
4 GeneArt® Gene Synthesis Services

5. A B
Wild type Optimized CREB1
x 2.8 References
Relative expression
mock
PP3
PP3
PP2
PP2
PP1
PP1
1. Fath S, Bauer AP, Liss M et al. (2011)
Multiparameter RNA and codon
60 optimization: a standardized tool to assess
CREB1 and enhance autologous mammalian gene
35
expression. PLoS One 6(3):e17596.
wild type optimized 2. Raab D, Graf M, Notka F et al. (2010) The
GeneOptimizer algorithm: using a sliding
SMARCD1 window approach to cope with the vast
x 1.8
sequence space in multiparameter DNA
Relative expression
sequence optimization. Syst Synth Biol
4(3):215–225.
60
SMARCD1
40
wild type optimized
JNK3
x 15.0
Relative expression
60
JNK3
40
wild type optimized
AQP5
x 9.0
Relative expression
60
AQP5
30
wild type optimized
IL-2
x only opt
Relative expression
20
IL-2
15
wild type optimized
Figure 2. Comparative expression analysis. Comparative expression analysis of wild type versus opti-
mized genes representing different protein classes. (A) Cell culture supernatants (immunomodulators,
IM) or cell lysates (all other protein classes) were analyzed by western blots using the α-Penta-His
antibody. One example from each protein class is shown. A cross-reactive 60 kD protein used to
standardize protein amounts is visible, including in the empty-vector negative controls (mock). Left:
molecular weight (kDa) markers, right: arrows indicating specific protein bands. (B) Relative expression
levels were derived by comparing mean expression (three independent transfections) of wild type or
optimized constructs, with wild type set to 1. The X-fold expression increase following gene optimization
is indicated for each protein (only opt = no detectable wild type expression).
To learn more and place your order, go to www.invitrogen.com/genesynthesis 5

6. Gene synthesis
gene synthesis has become a cost-effective, time- and resource-saving method for obtaining nearly any desired DnA
construct with 100% accuracy. It outperforms conventional molecular biology techniques in terms of time and cost, while
providing equivalent or better expression performance, and construct stability and quality. geneArt® gene synthesis tools go
beyond traditional synthesis and enable expression optimization and maximum performance.
Benefits
• Proprietary expression and mRNA stability optimization www Day 1
• Unlimited flexibility in gene and vector design Ordering until 3:00 pm (CET)
• Empirically proven expression increases G
A
• Ready-to-use constructs for expression and transfection
A
G A G
G
5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘ 5‘
Oligo synthesis overnight
• Easy online ordering
Quality
• All processes are ISO 9001:2008 quality certified Day 2
• Comprehensive quality documentation included Gene Assembler ® gene synthesis
platform
• Automated production processes
SuperSPEED Day 3
• Up to 1,200 bp in 5 business days (Figure 3) Cloning
• Up to 1,800 bp in 7 business days
• Emergency genes on request
Day 4
Performance A C G C A Sequencing quality control
• Project setup assistance and individual project support
• Maximum performance is available using the GeneOptimizer®
algorithm—the industry-preferred optimization algorithm Day 5
• Maximum production speed and worldwide delivery; capacity and Ready for shipment
reliability is supported by the world’s only industrial gene-processing
platform Figure 3. SuperSPEED gene synthesis service production
schedule.
Gene synthesis by GeneArt®
GeneOptimizer®
optimization algorithm
wild
type
GeneAssembler®
gene-synthesis platform
expression yield
Classic cloning
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Figure 4. GeneArt® gene synthesis and gene optimization is faster than classic cloning and can provide better results.
6 GeneArt® Gene Synthesis Services

7. Directed evolution
Directed evolution strategies are the most efficient method for creating proteins with improved or novel properties. The
directed evolution technologies from geneArt® synthesis help to evolve proteins in a goal-oriented, systematic process.
Site-directed mutagenesis 50 60 70 80
Introduce single or multiple mutations (substitutions, insertions, or
wild type GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT
deletions) into existing DNA sequences. F58A GVVPILVELDGDVNGHKASVSGEGEGDATYGKLTLKFICT
Benefits: fully sequence-verified clones, no unwanted backbone F59A GVVPILVELDGDVNGHKFAVSGEGEGDATYGKLTLKFICT
F60A GVVPILVELDGDVNGHKFSASGEGEGDATYGKLTLKFICT
mutants, and the fastest turnaround times. F61A GVVPILVELDGDVNGHKFSVAGEGEGDATYGKLTLKFICT
Applications: construction of fusion proteins, tagged proteins,
alternative splice forms, alanine scans, etc.
Site-saturation mutagenesis
Scanning a protein region by site-saturation mutagenesis identifies all 50 60 70 80
beneficial substitutions for enhanced function.
wild type GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT
Benefits: best cost efficiency with no structural data needed for protein F58A GVVPILVELDGDVNGHKASVSGEGEGDATYGKLTLKFICT
improvement. F58C GVVPILVELDGDVNGHKCSVSGEGEGDATYGKLTLKFICT
F58D GVVPILVELDGDVNGHKDSVSGEGEGDATYGKLTLKFICT
Applications: improvement of (industrial) proteins, alienation of proteins F58E GVVPILVELDGDVNGHKESVSGEGEGDATYGKLTLKFICT
from patented sequences, etc.
Combinatorial libraries
True rational design for defined randomization of selected sites only, 50 60 70 80
while providing maximum framework integrity.
wild type GVVPILVELDGDVNGHKXXVSGXGEXXATYGKLTLKFICT
Benefits: lowest ancillary mutation rates and highest diversities. peer A01 GVVPILVELDGDVNGHKRQVSGGGEGDATYGKLTLKFICT
Applications: construction of recombinant antibody libraries, promoter peer A02 GVVPILVELDGDVNGHKAGVSGEGEGDATYGKLTLKFICT
peer A03 GVVPILVELDGDVNGHKIYVSGLGEGDATYGKLTLKFICT
libraries, and combination of substitutions identified by site-directed peer A04 GVVPILVELDGDVNGHKLKVSGPGEGDATYGKLTLKFICT
mutagenesis.
Controlled randomization libraries
Substitute any amino acid in a gene with a defined probability. 50 60 70 80
Benefits: accurate fine-tuning of mutation rate, and randomization of wild type GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLXXICT
the entire open reading frame. peer A01 GVVPILVELDGDVNGHKFTVSGEGEGDATYGKLTLLKICT
peer A02 GVVPILVELDGDVNGHKTSVSGEGEDDATYGKLTLIGICT
Applications: affinity maturation of antibodies, improvement of industrial peer A03 GVVPILVELDGDVNGHKFSVSGAGEGDATYGKLTLQDICT
enzymes, modification of enantioselectivity of enzymes, etc. peer A04 GVVPILVELDGDVNGHKFSVSGEGEGYATDGKLTLLPICT
Truncation libraries
50 60 270 280
Create custom-defined populations of up to 40,000 in-frame truncated
constructs. wild type GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT
peer A01 VVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT
Benefits: Highest quality by avoiding out-of-frame mutations. peer A02 GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
Applications: solubility screen, minimal functional-size evaluation, peer A03 ILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
peer A04 DGDVNGHKFSVSGEGEGDATYGKLTLK
domain identification, inhibitory screenings, epitope mapping, etc.
To learn more and place your order, go to www.invitrogen.com/genesynthesis 7

8. Cell lines and proteins
Starting with only the nucleotide sequence, geneArt® services can provide purified protein within 30 business days. Protein
purification from transiently transfected mammalian cells assures correct folding and processing. Usage of optimized
genes for stable cell-line generation leads to production of high levels of active protein.
Benefits
• Seamless project processing—gene synthesis and protein purification from one source
• Speed—from gene to protein within 30 business days
Services
Expression analysis Optimized gene Genes to expression
in expression expression
Verification of your expression construct or vector verification
evaluation of the best variant of your protein
Protein production Genes to proteins
‘‘pilot‘‘
Genes to proteins
‘‘scale‘‘
Genes to proteins
‘‘guaranteed‘‘
Protein production in mammalian cells using Pilot production Customer–defined
culture volume
Guaranteed
amount
advanced transient transfection protocols
Cell line development Genes to cell lines
‘‘polyclonal‘‘
Genes to cell lines
‘‘clonal‘‘
Generation of cloned or uncloned stable cell lines Stable cell line
polyclonal
Stable cell line
clonal
Examples
• Improved expression reliability—optimized genes show expression of otherwise non-expressible proteins (Figure 4)
• Protein production by transient transfection—natural or approved artificial leader-peptides yield excellent secretion of engineered protein into the
culture supernatant (Figure 5)
• Improved transgene expression—productivity of antibodies from stable cell lines is improved with optimized expression constructs (Figure 6)
1 2 3
mock wild type optimized BSA standard ECD of
kDa 1 2 3 1 2 3 receptor tyrosine kinase
20 monoclonal antibody
IL-2
15
Figure 5. 10 mg of the extracellular domain of a receptor tyrosine kinase and a
monoclonal antibody (heavy and light chain) were purified from the supernatant of
transiently transfected HEK293/CHO cells via affinity tag/ProteinA.
IL-2
x only opt
500
Relative expression
protein concentration (µg/mL)
best producer
400
300
200
100
0
wild type optimized
wild type optimized
Figure 4. Three independent transfections of each wild type and optimized IL-2 gene Figure 6. Stable cell lines expressing different combinations of HC and LC of a human
were analyzed by western blot and densitometric analysis of the resulting bands. antibody were generated with wild type and optimized sequences and antibody pro-
duction yield was compared.
8 GeneArt® Gene Synthesis Services

9. Plasmid services
geneArt® plasmid DnA purification protocols help ensure consistent high quality for research applications and preclinical
studies. From vector construction to the production of plasmid DnA for preclinical trials, geneArt® plasmid services make
the development and execution of your project easy.
Gene synthesis
High-quality, scalable plasmid DNA for all applications
• Highly pure and homogeneous plasmid DNA
• Low levels of endotoxin (down to 0.01 EU/μg pDNA)
• Milligram to gram scale
• Fill-and-finish service
Subcloning
Applications
• Cell transfection
• Immunization studies
• Preclinical studies
• Toxicological studies
• DNA vaccine research
Plasmid production
Vector construction
Using expression-optimized, exchangeable genetic elements, GeneArt®
plasmid services manage individual vector design and complex subclon-
ing projects, delivering streamlined plasmid production and optimal
gene expression.
Project documentation Vector construction
A certificate of analysis (CoA) is provided with every plasmid order.
Premium documentation of all DIN ISO–certified production processes
can be provided on request.
Production documentation
To learn more and place your order, go to www.invitrogen.com/genesynthesis 9

10. 10 GeneArt® Gene Synthesis Services

11. To learn more and place your order, go to www.invitrogen.com/genesynthesis 11

12. For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
© 2011 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.
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