StarGate® offers rapid and highly efficient subcloning of an arbitrary gene – initially cloned into a Donor Vector - to simultaneously fuse it with many different genetic surroundings via transfer into Acceptor Vectors to generate Destination Vectors. The latter enable the efficient expression of your protein with various features (e.g. different tags and different promoters) in different hosts.
- Minimal modification of the gene of interest due to short combinatorial sites(4 bases only).
- Easy-to-handle subcloning procedure.
- Systematic screening of different elements (e.g. tags/promoters) in a variety of hosts.
- Inherent high level cloning efficiency due to a directed reaction(no equilibrium.
- Availability of a multitude of combinatorial sites for combinatorial cloning.
- Strep-tag® Expression Vectors
For expression in E. coli we are offering a large variety of pASK-IBA vectors allowing the expression of recombinant proteins carrying Strep-tag. Except for the antibiotic resistance genes (AmpR, CamR) the pASK-IBA vectors differ only between the XbaI and HindIII restriction sites.
Mammalian: pEXPR-IBA vectors are designed for high-level expression and purification of recombinant Strep-tag®/One-STrEP-tag and/or 6xHistidine-tag fusion proteins in mammalian cells. The vectors provide the same cloning strategy and are, thus, compatible with the corresponding bacterial pASK-IBA plasmids (see vectors for E. coli). As a consequence a PCR fragment can be cloned into pASK-IBA and its pEXPR-IBA equivalent in parallel (e.g. pASK-IBA3 and pEXPR-IBA3). The human cytomegalovirus (CMV) immediate early promoter provides strong expression in a wide range of mammalian cells. To prolong expression in transfected cells, the vector will replicate in cell lines that are latently infected with SV40 large T-antigen (e.g. COS7). In addition, the Neomycin resistance gene allows direct selection of stable cell lines.
Yeast: The vectors pKS1-ST and pKS2-ST combine two important features for protein expression:
- Inducible ADH2 promoter: The ADH2 promoter is induced by depletion of glucose in the medium. During early growth phase when glucose is abundant, the promoter is inactive, thereby preventing negative effects on yeast growth. When the cells reach the early stationary phase glucose is depleted from the medium and the ADH2 promoter is induced, producing high amounts of protein.
- KanMX cassette: Unlike other yeast expression vectors, the pKS-ST vectors feature a KanMX cassette, which allows selection with G418 in rich medium. The use of rich medium such as YPD will significantly enhance cell densities and protein yield.
Name Catalog # Type Species Applications Size Price pASK-IBA2C 2-1321-000 Expression Vector E. Coli 5 ug $232.00 pASK-IBA3C 2-1322-000 Expression Vector E. Coli 5 ug $232.00 pASK-IBA4C 2-1323-000 Expression Vector E. Coli 5 ug $232.00 pASK-IBA5C 2-1324-000 Expression Vector E. Coli 5 ug $232.00 pASK-IBA6C 2-1325-000 Expression Vector E. Coli Targeted Protein Expression 5 ug $232.00 pASK-IBA7C 2-1326-000 Expression Vector E. Coli Targeted Protein Expression 5 ug $232.00
- Strep-Express Vectors
Strep-Express vectors provide an easy and straight-forward entrance into the Strep-tag® technology. Strep-Express comprises Twin-Strep-tag® vectors for N- or C-terminal fusion, vectors for different expression hosts and the option for protein secretion.
In a first step the PCR fragment is generated using the respective forward (CF) and reverse (CR) primer extending the gene of interest (GOI) with the corresponding integration sites. This PCR product is in a next step integrated into the appropriate Acceptor Vector resulting in the final expression construct, the so called Destination Vector. The formation of the correct Destination Vector is monitored via blue-white screening on LBAgar plates containing X-Gal. The validated Destination Vector can be directly used for transformation or transfection of the expression host.
Name Catalog # Type Species Applications Size Price Forward and Reverse sequencing primers for pESG-IBA and pEXPR-IBA vectors 5-0000-124 Gene Transfer 1 nmol each $45.00 pASG-IBA102 5-4102-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA103 5-4103-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA104 5-4104-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA105 5-4105-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA123 5-4123-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA143 5-4143-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA144 5-4144-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA145 5-4145-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA162 5-4162-001 Expression Vector Gene Transfer 5 ug $268.00 pASG-IBA164 5-4164-001 Expression Vector Gene Transfer 5 ug $268.00