The pgex vectors have an expanded multiple cloning site mcs that contains six restriction sites. Pmalc5x vector 10 ug, 10 g, pmalc5x, n8108s, 101641058, each, login. Product components and storage conditions product size cat. The enzymes chosen are those which we believe to have been commercially available in june 1992. A series of sitespecific proteases complements the. This is a free resource for the scientific community that is compiled by addgene this page is informational only this vector is not available from addgene please contact the manufacturer for further details. The complete nucleotide sequence and list of the complete nucleotide sequence and list of restriction sites can be found at. The designed fusion gene strategy allows the purification of soluble and insoluble recombinant proteins to homogeneity with singlestep affinity. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 phone 6082744330 fax 6082772516 printed in usa. Polylinkers of pgex4t1, pmalc2, and a pet vector were replaced with the.
Map of the glutathione stransferase fusion vectors showing. What are the steps for cloning a gene fragment in the pgex. The pcmvscript vector is derived from a highcopynumber pucbased plasmid and is designed to allow protein expression in mammalian systems. 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. Map of the glutathione stransferase fusion vectors showing reading frames and main features. Figure 2 circular map and features for the pfbneo retroviral vector. Home resources plasmid files pgex vectors ge healthcare pgex vectors ge healthcare. Some of the vectors contain the 35s promoter upstream of the cloning site while others have none.
A cterminal v5 epitope and polyhistidine 6xhis tag allow for detection using an antiv5 antibody and an antihis antibody respectively. 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. T, pgex6p1, pgex4t1, and pgex5x1 can directly accept and. Mbp fusions are also easily affinity purified on amylose columns. Note that the sequence is numbered by the pbr322 convention, so the t7 expression region is reversed on the circular map. Please contact your local pharmacia subsidiary if a discrepancy is identified. Bacterial vector for expressing fusion proteins with a thrombin site. Restriction sites for pgex4t1 the following restriction site table was compiled using dnasis software for sequence analysis. This vector can be used in combination with pacycduet1 cat. The pmalpiii vector is a derivative of pmalp2 in which the leader sequence of maltose binding protein mbp, male has been. The same multiple cloning sites mcs in each vector ensure easy transfer of inserts. Sca i 1875 ori pgemt vector 3003bp ampr apa i aat ii sph i bstz i nco i sac ii spe i not i bstz i pst i sal i nde i sac i bstx i nsi i t7 xmn i 1994 nae i 2695 lacz f1 ori 1 start 14 20 26 31 37 46 55 62 62 73 75 82 94 103 112 126 sp6 t t sca i 1890 ori pgemt easy vector 3018bp. This products resides on a fisher scientific gsa or va contract. The n or cter minal tags are automatically fused subsequent to.
Configurations available with the synthetic firefly luc2 photinus pyralis and renilla hrluc renilla reniformis genes, which have been codon optimized for more efficient expression in mammalian cells. Promega corporation2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 telephone 6082744330 fax 6082772601 printed in usa. T 27480501 pgex6p1 27459701 bamh i ecor i sma i sal i xho i not i prescission protease leu glu val leu phe gln gly pro leu gly ser pro glu phe pro gly arg leu glu arg pro his. This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Find msds or sds, a coa, data sheets and more information. Map of the gst vectors showing the reading frames and main features. This is a free resource for the scientific community that is compiled by addgene. The n or cter minal tags are automatically fused subsequent to the lr reaction.
Even though stop codons in all three frames are not depicted in this map, all thirteen vectors have stop codons in all three frames downstream from the multiple cloning site. T, pgex6p1, pgex4t1, and pgex5x1 can directly accept and express cdna inserts isolated from. Data file 28962284 aa gst gene fusion system glutathione. Thrombin tac promoter lac o rbs bamh i ecor i sma i sal i xho i not i a m p r m c s i a c i g s t p b r 3 2 2 o r i a m p r pgex 4t1 5. These combined dna sequence and map files can be opened with snapgene or the free snapgene viewer. Mammalian expression is driven by the human cytomegalovirus cmv. A differentially regulated ap2erf transcription factor. All thirteen vectors have stop codons in all three frames downstream from the multiple cloning site not depicted in this map. Data file 28962284 aa gst gene fusion system glutathione s. An equimolar amount of attb pcr product or linearized attb expression clone and the donor vector. T 27480501 pgex6p1 27459701 bamh i ecor i sma i sal i xho i not i prescission protease leu glu val leu phe gln gly pro. The sale of pdonr p4p1r multisite gateway threefragment vector construction kit.
The adoptive transfer of tumorreactive t cells has been shown to mediate the regression of tumors. Both the reporter genes and the vector backbone, including the bla. The vector pgex4t1 was not tested with each enzyme and therefore the accuracy of the table cannot be guaranteed. I have designed the primers with the ecor1 and sal1 enzyme sites at 5. Gst gene fusion system pennsylvania state university. Locus exported 4969 bp dsdna circular syn 031220 definition bacterial vector for expressing fusion proteins with a thrombin site. What are the steps for cloning a gene fragment in the pgex 4t1 vector. Map of the glutathione stransferase fusion vectors showing the reading frames and main features. Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Thrombin tac promoter lac o rbs bamh i ecor i sma i sal i xho i not i a m p r m c s i a c i g s t p b r 3 2 2 o r i a m p r pgex4t1 5. Please note that enzymes with nonpalindromic recognition sequences are listed twice. The expanded mcs facilitates the unidirectional cloning of cdna inserts obtained from libraries constructed using many available lambda vectors. A novel expression vector pgex5t was constructed which directs the synthesis of a fusion protein with a histidinehexapeptide and glutathionestransferase at its nterminus and the recombinant protein at its cterminus inescherichia coli. A differentially regulated ap2erf transcription factor gene.
Description continued the pgl4 vector family includes. New vectors for high level expression of recombinant. Adoptive immunotherapy using tcr genemodified t cells is a promising strategy for producing tumor. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. The resulting vector, pgexkg, produced a fusion protein which was readily cleaved by thrombin, but the resulting protein had several glycine residues at its amino terminus. The pgem t and pgem t easy vector systems have been optimized using a 1. To support our community, tair access limits have been lifted until may 31. If initial experiments with your pcr product are suboptimal, ratio optimization may be necessary. The designed fusion gene strategy allows the purification of soluble and insoluble recombinant proteins to homogeneity with.
Collectively, the pgex vectors provide all three translational reading frames beginning with the ecori restriction site. Orfs inserted into mcs1 can be sequenced using the. Basic vectors with no promoter that contain a multiple cloning region for cloning a promoter of choice. Donor vector to use in the reaction for optimal efficiency, we recommend using the following amounts of attb pcr product or linearized b expression clone and donor vector in a 20 l bp recombination reaction. Bacterial vector for expressing gst fusion proteins with a factor xa site.
The usefulness of this vector is re flected in the fact that we have had over 300 requests for the pgexkg plasmid. The glutathione stransferase gst gene fusion system is an integrated system for the expression, purification and detection of fusion proteins produced in e. In order to produce recombinant crmapkk1 protein, the corresponding gene was pcr. The pgex vectors are designed for inducible, highlevel intracellular expression of genes or gene fragments. See appendix 2 control regions for pgex vectors for the control regions of the vectors. Map of the glutathione stransferase fusion vectors showing the reading frames. Eukaryotic expression vector containing the complete coding sequence of l. The pmalpiii vector is a derivative of pmalp2 in which the leader sequence of. The vector has the pbr322 lowcopy origin of replication for minimized basal expression level. All pgwb vectors confer resistance to both kanamycin and hygromycin in plant. New vectors for high level expression of recombinant proteins.