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Trusted by Leading Research & Pharma Institutions

Gateway Cloning Services

Accelerate your molecular cloning with our proprietary Gateway recombination technology. Transfer genes seamlessly between vectors without restriction enzymes or ligases, achieving >95% cloning efficiency in just one hour.

>95% Efficiency
1hr Reaction
No Enzymes Needed
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Trusted by leading research and pharmaceutical institutions

Harvard
Pfizer
MIT
Roche
Stanford
Novartis

Why Choose Us

Proprietary recombination enzymes
Universal vector compatibility
100% sequence verified
Multi-fragment assembly

BP Reaction

attB × attP → attL Entry Clone

LR Reaction

attL × attR → Expression Clone

MultiSite Assembly

Up to 4 fragments in one reaction

Cloning Efficiency
>95%
Overview Technology Specifications Workflow Applications FAQ
Service Overview

Streamlined Gateway Recombination Cloning

Our proprietary Gateway cloning platform enables seamless gene transfer between vectors with unmatched efficiency and reliability.

Fast Recombinational Cloning

Our proprietary enzyme mixes catalyze site-specific recombination between att sites in just one hour at room temperature. No restriction enzymes, no ligase, no gel purification required.

  • 1-hour reaction at 25°C
  • >95% cloning efficiency
  • Directional and in-frame fusion

Universal Vector Compatibility

Transfer your gene of interest from entry clones to hundreds of Gateway destination vectors for expression in bacterial, yeast, insect, and mammalian systems.

  • 100+ destination vectors available
  • Multiple expression systems
  • Flexible tag options

Reversible Reactions

Both BP and LR reactions can be reversed for fragment recovery or reuse.

Multi-Site Assembly

Clone 2-4 DNA fragments simultaneously into a single vector in one reaction.

Sequence Verified

Every clone is Sanger sequenced to guarantee 100% accuracy.

Ready to Simplify Your Cloning?

Get expert support for your Gateway cloning project.

Technology Platform

Proprietary Recombination System

Based on λ integrase site-specific recombination for highly efficient and directional cloning.

BP Clonase II Mix

Catalyzes attB × attP recombination to generate attL-flanked entry clones. Highly efficient and reliable.

>1500 colonies 1 hour

LR Clonase II Plus

Transfers genes from entry clones to destination vectors. Generates >5000 colonies per reaction.

>5000 colonies High efficiency

MultiSite Pro Plus

Assemble 2-4 DNA fragments simultaneously in a single reaction with precise orientation.

2-4 fragments Directional

Quality Control

SEQ Sanger sequencing verification
COA Certificate of Analysis included
ccdB Negative selection marker

Expression Systems

BAC E. coli expression
YEAST Saccharomyces cerevisiae
MAM Mammalian cells (HEK293, CHO)
Specifications

Flexible Options for Diverse Needs

Comprehensive specifications to meet your research requirements.

Parameter Entry Clone Construction Expression Clone MultiSite Assembly
Insert Size 100 bp - 11 kb Up to 15 kb 2-4 fragments
Reaction Time 1 hour (25°C) 1 hour (25°C) 16-18 hours
Cloning Efficiency >95% >95% 70-90%
Verification Sanger sequencing Sanger + restriction Colony PCR + seq
Selection Kanamycin + ccdB Ampicillin + ccdB Multiple markers
Delivery Format Plasmid DNA + glycerol Plasmid DNA + COA Validated clones
Workflow

Simple 5-Step Process

Our streamlined workflow ensures efficiency and reliability at every stage.

1

Design

Submit sequences and vector specs

2

BP Reaction

Entry clone construction

3

LR Reaction

Expression clone generation

4

QC

Sanger sequencing verification

5

Delivery

Plasmid + COA report

Applications

Diverse Applications Across Biotechnology

Our Gateway cloning services support research and development in multiple fields.

Protein Expression Screening

Rapidly transfer your gene of interest into multiple destination vectors to find the optimal expression system. Test different tags, promoters, and host cells in parallel.

  • Bacterial, yeast, insect, and mammalian expression
  • N-terminal and C-terminal fusion tags
  • GFP, His-tag, Flag-tag, GST options
  • Codon optimization available
100+
Expression vectors available

Functional Genomics & cDNA Libraries

Clone entire cDNA libraries into entry clone collections for high-throughput functional analysis. Transfer clones to diverse assay vectors for systematic studies.

  • Full-length ORF cloning
  • Yeast two-hybrid compatible
  • High-throughput formats
  • 96-well plate options
HTP
High-throughput compatible

Multi-Site Gateway Assembly

Combine 2-4 DNA fragments (promoter, ORF, tag) simultaneously into a single vector in one reaction. Maintain precise orientation and reading frame.

  • 2, 3, or 4 fragment assembly
  • Precise order and orientation
  • No restriction enzymes needed
  • Single reaction simplicity
4
Fragments in one reaction
Testimonials

What Our Clients Say

Trusted by researchers worldwide for quality and reliability.

"The cloning efficiency is remarkable. We've switched from traditional restriction cloning to Gateway and our project timelines have shortened significantly. The technical support is excellent."

M
Senior Scientist
Biotechnology Company

"Gateway cloning has been essential for our high-throughput protein expression screen. The ability to transfer ORFs to multiple vectors in parallel has accelerated our discovery pipeline."

J
Research Director
Academic Research Institution

"We use MultiSite Gateway for assembling complex multi-component constructs. The efficiency and directionality are excellent. Sequencing verification gives us confidence in every clone."

K
Principal Investigator
Pharmaceutical Company
Scientific Literature

Scientific Foundation

Our platform is backed by peer-reviewed research.

89 Citations

Structure-specific DNA recombination sites: Design, validation, and machine learning–based refinement

Nivina A, Grieb MS, Loot C, et al. Science Advances. 2020.

Development of structure-specific recombination technology based on integrase systems with minimal sequence constraints.

View DOI
156 Citations

CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering

Vo PLH, Ronda C, Klompe SE, et al. Nature Biotechnology. 2020.

INTEGRATE system achieving ~100% efficiency for precise, marker-free DNA integration up to 10 kb.

View DOI
12 Citations

Fusion-PCR generates attL recombination site adaptors and allows Rapid One-Step Gateway (ROG) cloning

Yang D, Liu C, et al. Biochimie. 2020.

Rapid One-Step Gateway cloning method eliminating BP reaction step, saving time and cost.

View DOI
245 Citations

Gateway Recombinational Cloning

Reece-Hoyes JS, Walhout AJM. Cold Spring Harbor Protocols. 2018.

Comprehensive protocol guide for Gateway cloning system based on λ integrase recombination.

View DOI
78 Citations

A non-viral genome editing platform for site-specific insertion of large transgenes

Multiple authors. Nucleic Acids Research. 2020.

Lambda integrase-based platform for efficient large transgene (>10kb) insertion in human cells.

View DOI
FAQ

Frequently Asked Questions

Find answers to common questions about our Gateway cloning service.

Gateway cloning is a site-specific recombination method based on λ integrase technology. It uses two reactions: BP reaction (attB × attP → attL entry clone) and LR reaction (attL × attR → expression clone). Unlike traditional restriction cloning, it doesn't require compatible restriction sites and maintains direction and reading frame.
Key advantages include: >95% cloning efficiency, 1-hour reactions at room temperature, no need for restriction enzymes or ligase, directional cloning that maintains reading frame, and the ability to transfer genes to hundreds of different vectors without re-cloning. The system is also reversible, allowing fragment recovery.
Standard Gateway cloning works well with inserts from 100 bp to 11 kb. Larger inserts up to 15 kb can be cloned with extended reaction times. For MultiSite assembly, 2-4 fragments of varying sizes can be combined in a single reaction.
We offer Gateway destination vectors for bacterial (E. coli), yeast (S. cerevisiae), insect (Sf9, High Five), and mammalian (HEK293, CHO) expression systems. Vectors include various promoters (constitutive, inducible), tags (His, GST, Flag, GFP, etc.), and selection markers.
Yes, we can work with your existing Gateway-adapted vectors (containing attR sites and ccdB cassette). If your vector is not Gateway-compatible, we offer a vector conversion service to insert the necessary recombination sites.
All clones undergo Sanger sequencing verification across recombination sites and the entire insert. Restriction digest analysis confirms clone identity. Final deliverables include purified plasmid DNA, bacterial glycerol stock, Certificate of Analysis, and sequencing data.

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Expert consultation

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