Pichia pastoris expression vector with methanol inducible promoter AOX1 and the alpha factor secretion peptide. Also has a histidine synthesis selection marker.
Product Name: pSF-AOX1-A-factor-SEAP-HIS4
Product Code: OG3583
Size (bp): 8693 bp
Bacterial Antibiotic Selection: KanR
Origin and Compatibility: pUC high copy derived from pBR322
Bacterial Copy Number: 500-700 per cell
This vector encodes the SEAP polypeptide as a positive control for secreted protein expression. This vector is designed for the expression of a gene of interest (GOI) in the methylotrophic yeast, Pichia pastoris. Once the GOI has been cloned into the MCS, the plasmid is then linearised and transformed into an appropriate P. pastoris strain by electroporation. The expression construct becomes integrated into the genome. Expression of the GOI is induced by addition of methanol to the culture medium.The vector contains a HIS4 marker for nutritional selection of integrants in a HIS4 P. pastoris strain.This vector enables secretion of your GOI using the upstream alpha-factor secretion signal.Promoter Expression Level:
This promoter is inducible by methanol and exhibits high expression under these conditions. In the absence of methanol the promoter is tightly regulated and shows only basal levels of expression.About the Peptide Tag:
This plasmid contains an full length Alpha Factor (A-FactorFL) secretory peptide to allow proteins to be exported from the cytosol. Alpha factor is an endogenous yeast protein that is naturally secreted into the supernatant. The wild-type Alpha Factor protein contains a series of repeating protein sequences that are cleaved out by cellular proteases that recognise a specific site. This produces a series of small peptides that contain the mature alpha peptide sequence. This plasmid contain the wild-type signal peptide from the full length Alpha factor gene but also the coding sequence that leads up to the first endogenous protease cleavage site. The addition of these sequences has been demonstrated to provide more consistent secretion of proteins in comparison to using just the signal peptide alone. All of the wild-type protein sequences will be removed during protein maturation and export by endogenous proteases.
This plasmid contains the poly-adenylation signal from the Alcohol Oxidase 1 Promoter (AOX1) gene from Pichia pastoris to terminate transcription.
This plasmid is not designed to clone in a gene, instead it is designed for testing secretion in Pichia pastoris.Multiple cloning site notes:
This product is part of our SnapFast™ plasmid range, for more information on the intellectual property status of this plasmid and the terms of our licences please click here.