Schematic diagram describing the Gateway cloning
(Barbara Meissner,et al.,2011)
Gateway Cloning Service is a specialized service offered by our company that allows for efficient and precise transfer of DNA fragments into a variety of expression vectors. This technique is widely used in molecular biology research and enables seamless assembly of DNA constructs. Whether you need to clone small or large DNA fragments, our Gateway Cloning Service provides a reliable and versatile solution.
Our Gateway Cloning Service follows a systematic process to ensure a seamless and efficient cloning experience. Here is an overview of the steps involved:
For more information about our Gateway Cloning Service and to discuss your project requirements, please contact our team by submitting your contacts, We are here to assist you every step of the way.
Multisite Gateway-mediated Cas9/gRNA expression system (GCas method). (A) sgRNA module vectors harboring the att sites. U6, mammalian U6 type III RNA polymerase III promoter; H1, H1 RNA polymerase III promoter; BbsI, a single sgRNA cloning site mediated by BbsI digestion; BsmBI, a dual sgRNA cloning site mediated by BsmBI digestion; Scaff, scaffold sequence of sgRNA followed by TTTTTT (polyT terminal signal of RNA polymerase III); C4s, a Csy4-mediated RNA cleavage site. See Figure E in S1 Supporting Information and the corresponding legend for the cloning of sgRNA(s) into the sgRNA module vectors. (B) Promoter vectors harboring attL5 and attR3. CMV, cytomegalovirus promoter; CAG, cytomegalovirus enhancer fused to the chicken beta-actin promoter and introns of chicken beta-actin and rabbit beta-globin; EF1α, Elongation factor 1-alpha promoter. (C−D) Vectors harboring Cas9 alone or Cas9/EGFP harboring attL3 and attL2. 2As are illustrated as right-pointing triangles. Cas9, spCas9; 2A, Thosea asigna virus 2A peptide sequence (T2A); Csy4, Csy4 RNase of Pseudomonas aeruginosa; FokI, dimerization-dependent wild-type FokI nuclease-domain; Cas9n, mutated Cas9 nickase (D10A); dCas9, catalytically inactive Cas9. (E) Destination vectors for the GCas method. The CmR and ccdB gene are located between attR1 and attR2 in each destination vector (not described) for the positive/negative selection of transformants harboring the vector itself or the desired Cas9/gRNA expression vector. Gray arrows and boxes in CSIV-DEST indicate regulatory sequences for lentivirus production such as 5'LTR (5'-side long terminal repeat), C (Packaging signal), cPPT (central polypurine tract), WPRE (woodchuck hepatitis virus posttranscriptional regulatory element) and 3'LTR (3'-side long terminal repeat). 5'ITR and 3'ITR, 5'-side and 3'-side of Piggybac inverted terminal repeats. The entire sequences of the vectors described are attached in the S1 sequence. (F) Other module vectors for the GCas method. pA, simian virus 40 polyadenylation signal sequence.——(S Yoshimatsu,2019)
Here are some frequently asked questions about our Gateway Cloning Service:
Gateway Cloning is compatible with a wide range of expression vectors, including plasmids and viral vectors. Our experts can guide you in selecting the most suitable vector for your specific application.
The turnaround time for Gateway Cloning can vary depending on the complexity of the project. During the consultation phase, our team will provide you with an estimated timeline based on the specific requirements of your project.
Absolutely! Gateway Cloning is suitable for cloning both small and large DNA fragments. Our experts have the expertise and resources to handle a wide range of fragment sizes.
Yes, we provide comprehensive services for downstream applications. In addition to Gateway Cloning, we offer services such as protein expression and purification. Our goal is to provide a holistic solution to meet your research needs.
Getting started with our Gateway Cloning Service is simple. Just contact our team to discuss your project requirements, and we will guide you through the process. Our experts are here to assist you every step of the way.
Please note that all services are for research use only. Not intended for any clinical use.
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