pET E. coli T7 Expression Vectors. The pET System is the most powerful system for the cloning and expression of recombinant proteins in E. coli. Driven by the strong bacteriophage T7 promoter and translation signals, Novagen's® pET System has been used to express thousands of different proteins in host cells expressing T7 polymerase. pGEX-6P-1 Sequence and Map pGEX-6P-1 Bacterial vector for expressing GST fusion proteins with a PreScission protease site. For other reading frames, use pGEX-6P-2 or pGEX-6P-3. Sequence Author: GE Healthcare Open in SnapGene Try SnapGene for Free Download Plasmid | Download SnapGene Viewer pGEM®-T Easy Vector Systems. PCR cloning vectors with 3 options for insert excision. A1360, A1380. pCMVT n T™ and pT n T™ Vectors. Convenient expression of cloned genes in vitro or in vivo. pCMVT n T™ Vector contains a CMV enhancer/promoter region, which enables strong constitutive expression in many cell types, while RNA phage promoters enable RNA synthesis in vitro. View 242 Lab Manual - Module 2 - Molecular Biology.pdf from BIOL 24200 at Purdue University. BIOL 24200 Laboratory Manual Module 2: Molecular Biology Spring 2020 Instructor: Dr. Olga Yurchenko Table. Study Resources. Main Menu; The 'Vector' DNA is a pGEX plasmid, The pGEX vectors have an expanded multiple cloning site (MCS) that contains six restriction sites. The expanded MCS facilitates the unidirectional cloning of cDNA inserts obtained from libraries constructed using many available lambda vectors. pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3 each encode the recognition sequence for site-specific cleavage by PreScission Protease between the GST domain and Overview. The pET vector system is a powerful and widely used system for expressing recombinant proteins in E. coli. The gene of interest is cloned into the pET vector under the control of the strong bacteriophage T7 transcription and translation regulatory system. Activation of expression is achieved by providing T7 RNA polymerase within the cell. Technical Manual#TM016.) Use the T7 Promoter Primer or the pUC/M13 Forward Primer to sequence ssDNA produced by the pGEM®-T and pGEM®-T Easy Vectors. Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA ·Toll Free in USA 800-356-9526 ·Telephone 608-274-4330 ·Fax 608-277-2601 · promega.com Part# TM042 Printed in USA. For manual purification of sample volumes up to 600 µl use GST SpinTrap™ microspin columns or GST MultiTrap™ 4B 96-well plates. GST vector products Code No. pGEX-4T-1 (25 µg) 28-9545-49 pGEX-4T-2 (25 µg) 28-9545-50 pGEX-4T-3 (25 µg) 28-9545-52 pGEX-5X-2 (25 µg) 28-9545-54 Download scientific diagram | 2 Expression vector pGEX-2T (GST Gene Fusion System, laboratory manual, 3 rd edition, Amersham Biosciences). from publication: Kinetic Characterization of Drosophila pGEX-4T-1 (27-4580-01) Thrombin Stop codons sail Xhol EcoRl Sma I Not I BarnH I (4869) Btgl (4761) Bsu361 8fuAi - 'J Created With SnapGene. BspM1 (62) Can be used for tag cleavage when the PreScission Protease recognition sequence occurs between the tag sequence and the protein of interest. (e.g., the GST tag from proteins expressed using the pGEX-6P vector). Bovine thrombin: 37 000: 500 units: One unit cleaves > 90% of 100 µg of a test GST-tagged protein when incubated in 1× PBS at 22°C pGEX-6P-1 5.0kb. tae promoter Iac operator CST tag 207aa GGT G ACA ATT AAT CAT CGG CTC GTA TAA GTG GAA TOA GCG GAT AAC AAT rrc ACA CAG GAA ACA GTA rrc ATG TCC CCT ATA C,ST tag BamH 1 Smal Xholl Not