Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Each order of pet dna also includes an induction control strain supplied as a glycerol stock. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circular map. Twopromoter vector is highly efficient for overproduction of protein. Dec, 2005 therefore, the petm20eyfp vector was cultured using the degradationresistant carbenicillin instead of ampicillin and either transformed in plyss cells or cultured in the presence of glycerol to prevent expression leakage. We successfully constructed the recombinant plasmid pet30a ega31eg95 and expressed the recombinant protein ega31eg95. Can anybody suggest a vectors for cloning a gene between ncoi and hindiii site with histag at nterminal. The pet30 eklic vector is prepared for rapid, directional cloning of pcrampli. The pet32 vectors are sold under patent license from. Current information on the pet system may be obtained on line in a pdf file.
This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Bl21 competent cells 3 introduction host strains and genotypes for the escherichia coli strains, the genes listed in the table below signify that the bacterium carries a mutant allele. A repository of over 200,000 plasmids including protein structure initiative protein expression plasmids and vectors, over 75,000 human plasmids, and whole genome collections from many organisms. The pet system also uniquely provides six possible. The f1 origin is oriented so that infection with helper phage will produce virions containing singlestranded dna that corresponds to the coding strand. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circular. Andrew lees lab contains the insert hptp1e pdz2 and is published in biochemistry. Construction and identification of the recombinant plasmid. In the champion pet sumo protein expression system, you will clone and express your gene of interest as a fusion to sumo. The pet vectors are supplied as purified plasmid dna 10 g.
The positive clone was the empty recombinant vector. Popculture quick screen for expression level, activity, and solubility 25 b. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is re versed on the circular map. Our plasmid vectors enable you to choose the perfect combination of. Can anybody suggest a vectors for cloning a gene between. Sumo is the saccharomyces cerevisiae smt3 protein which is an 11 kda homolog of the mammalian sumo1 protein saitoh et al. The f1 origin is oriented so that infection with helper phage will produce virions. The pet30a vector was used for producing the target proteins without a tag fig. Terms and conditions 20042018 harvard medical school plasmid was created and is maintained by the dfhcc dna resource core at harvard medical schooldfhcc dna resource core at harvard medical school. The map, notes, and annotations on this page and in the sequence map file are ed material. We continue to work to improve your shopping experience and your feedback regarding this content is very important to us.
Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circle map. The cloningexpression region of the coding strand transcribed by t7 rna polymerase is shown below. A 0502 the pet44 vectors are designed for cloning and highlevel expression of peptide sequences fused with the 495 aa nustag protein. This is a free resource for the scientific community that is compiled by addgene. This allows the nterminus of the target protein to be fused to the intein tag. The pet vectors 4 vector characteristics and cloning strategy 4 ligationindependent cloning lic of. The f1 origin is oriented so that infection with helper phage will. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the. We seek to understand mechanisms of biology at the level of structures, interactions, and conformational changes of protein molecules and their interacting partners. Terms and conditions 20042018 harvard medical school plasmid was created and is maintained by the dfhcc dna resource core at harvard medical schooldfhcc dna resource. Terms and conditions 20042018 harvard medical school plasmid was created and is maintained by the dfhcc dna resource core at harvard medical schooldfhcc dna resource core at. Addgenes blog provides a platform for researchers to share information.
Bacterial vector for expressing thioredoxin fusion proteins with an enterokinase site. It therefore lacks the ribosome binding site and atg start codon present on the pet translation vectors. By simply adding a promoter sequence in front of the second target gene on a. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the vector map. Unique sites except for the two bstx i sites are shown on the circle map. Characterization of homodimer interfaces with cross. Embl is an intergovernmental organisation, consisting of more than 25 member states, associate and prospect members. We study multiple biological pathways and questions using a variety of approaches, especially electron cryomicroscopy cryoem, xray crystallography, and biochemistry. Vectors containing the histag sequence are licensed under u. Stabilize a toxic gene in ampr pet vectors during induction. As a result, the yields of the soluble proteins were dramatically increased fig. Note that the sequence is numbered by the pbr322 convention, so the t7 expression. Please use the form below to provide feedback related to. The pet32 and pet48 vectors are sold under patent license from genetics.
How to consider tags in vectors to help with protein purification when choosing restriction sites. Bacterial vector for expression of nterminally stagged proteins with an enterokinase site. Find msds or sds, a coa, data sheets and more information. Pod shattering resistance associated with domestication is. Cloning sites are shown on the maps provided with your pet vector kit. Dec 09, 2016 vector and host combinations affect expression levels 12.
Sgc oxford seeks to generate high quality reagents to accelerate the discovery of new medicines and place them into the public domain without restriction. How to consider tags in vectors to help with protein. A new vector for highthroughput, ligationindependent cloning encoding a tobacco etch virus protease cleavage site. Supplementary tables 15 and supplementary references pdf 10 kb rights and permissions. The vector map has been adapted with permission from tubb et. Using 3d structures of human proteins and the reagents generated as part of the structure determination process as well as the chemical probes identified, we work with organisations within oxford, the uk and the rest of the world to further. Smt3, hereby referred to as sumo, is a member of a. Take a look on your vector map, histaq is a part of your vector, upstream or downstream or. It contains the pmb1 origin of replication from pbr322 and is maintained at a similar copy number to pbr322. Addgene is a nonprofit plasmid repository facilitating life science research. To my knowledge the only suitable plasmid in the pet serie is the pet30a but it will.
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