PiggyBac Dual Promoter vector哺乳动物细胞双启动子转座子表达质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

PiggyBac Dual Promoter vector哺乳动物细胞双启动子转座子表达质粒载体

PiggyBac Dual promoter Information Promoter: CMV promoter Replicator: pUC ori, F1 ori Terminator: SV40 poly (A) signal Plasmid classification: lactation series plasmids; lactation editing plasmids; lactation transposing plasmids Plasmid size: 7258bp Prokaryotic resistance: ampicillin Amp (100 u g/ml) Screening markers: green fluorescent protein CopGFP, purinamycin Puro (10ug/ml) Cloned strains of Escherichia coli, DH5 A and other Escherichia coli Culture conditions: 37 centigrade, aerobic LB Expression host: mammalian cells such as 293T Culture conditions: 37 C, 5%CO2 Induction mode: no induction, instantaneous expression 5'sequencing primers: CMV-F (CGCAAATGGGCGGTAGGCGTG) 3'sequencing primers: Sv40-polyA-R (GAAATTTGTGATGCTATTGC)   Description   PiggyBac Dual promoter (PB513B-1) is a plasmid for mammalian cell transposition experiment, located at the downstream of CMV promoter, MCS, so that the target gene or microRNA is more convenient to clone. The downstream EF-1 alpha core promoter starts the expression of GFP. The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors and chromosomes via a "cut and paste" mechanism. During transposition, the PB transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes. The unique features of piggyBac transposons are that there is NO Cargo Limit and it is also Reversible. Genomes containing an inserted piggyBac vector can be transiently re-transfected with the PB transposase expression vector. The PB transposase will remove the transposons from the genome, footprint-free.The Super PiggyBac transposase transient expression vector and PB513B-1 were co-transfected into HeLa cells and puromycin selection applied for 10 days (10ug/ml). Cells efficiently transposed were Puro resistant and GFP positive.

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