Home > Protein Production & Assays > Cloning & Transfection > Cloning Strategies > Expression Vectors - Overview - Cloning

Expression Vectors at a glance

Overview on available StarGate expression vectors

StarGate Acceptor Vector collection

The StarGate Acceptor Vectors represent expression vectors providing different expression features like promoters, tags and signal sequences.

Acceptor Vectors for 4 different hosts and 5 different tags are currently available:

Host Promoter Tags
E. coli
Tet
T7*
Strep-tag®II,
Twin-Strep-tag®,
6xHis-tag,
FLAG-tag,
GST-tag with PreScission™ (PSC) site
Mammalia CMV Strep-tag®II,
Twin-Strep-tag®,
6xHis-tag,
FLAG-tag,
GST-tag with PreScission™ (PSC) site
Yeast CUP1 Strep-tag®II,
Twin-Strep-tag®,
6xHis-tag,
FLAG-tag,
GST-tag with PreScission™ (PSC) site
Insect cells (Baculovirus) Polyhedrin Strep-tag®II,
Twin-Strep-tag®,
6xHis-tag,
FLAG-tag,
GST-tag with PreScission™ (PSC) site

Expression vectors categorized by the fusion tag

Vectors without fusion tag
Twin-Strep-tag® vectors
Strep-tag® vectors

Classic cloning vectors for E. coli and mammalia

Cloning vectors for expression in E. coli and mammalian cells that provide different protease cleavage sites or chloramphenicol resistance.

IBA Expression Vectors:

E. coli

For recombinant protein expression in E. coli, vectors with the Tet promoter (pASK-IBA) are available. The choice of the best expression vector depends on the characteristics of the protein. Due to the identical multiple cloning sites different vector backbones can be easily tested. pASK-IBA vectors are available with different protease cleavage sites (Tev, Factor Xa, Thrombin and Enterokinase) or with chloramphenicol resistance gene.

Mammalia

Expression in mammalian cells can be achieved with the CMV promoter based expression vectors (pEXPR-IBA) for high protein yields in a wide range of mammalian cells. These vectors provide a N-terminal Strep-tag®II followed by a special protease cleavage site for removal of the tag after purification.

other vectors: His-, GST-, Flag-tag

Expression vectors categorized by expression host

E. coli

pASG-IBA vectors

  • High-level expression in E. coli, also for toxic proteins
  • Tightly regulated expression due to anhydrotetracycline (AHT) inducible tetA promoter/operator
  • Option for periplasmic expression due to ompA signal sequence
  • No catabolite repression - no influence of medium components
  • Not influenced by the genetic background – wide choice of E. coli expression strains
  • Inexpensive induction with AHT.

pPSG-IBA vectors

  • High-level expression in E. coli by bacteriophage T7 promoter
  • High-level transcription by T7 RNA polymerase in BL21 strains
  • High-level expression of non-toxic proteins
  • Induction by IPTG
  • Suitable for in vitro transcription/translation 
Mammalia

pDSG-IBA vectors

  • Optimized for MEXi, Mammalian Expression IBA, and are also suitable for other mammalian cell cultures
  • High level constitutive expression in mammalian cells by the CMV promoter
  • Extrachromosomal replication due to Epstein Barr Virus replication origin (oriP) - requires chromosomal expression of the EBNA-1 gene, e.g. like in MEXi 293E cells
  • ColEl ori and ampicillin resistance gene support propagation in E. coli
  • BM40 option for secretion of protein into the medium

pESG-IBA vectors

  • High-level constitutive expression in mammalian cells by the CMV promoter
  • Neomycin resistance for generation of stable cell lines
  • ColEl ori and ampicillin resistance gene support propagation in E. coli
  • BM40 option for secretion of protein into the medium

pCSG-IBA vectors

  • High-level constitutive expression in mammalian cells by the CMV promoter
  • extrachromosomal replication due to Epstein Barr Virus replication origin (oriP) and EBNA-1
  • prolonged expression of the inserted GOI under G418 selection without the need for making stable cell lines
  • ColEl ori and ampicillin resistance gene support propagation in E. coli
  • BM40 option for secretion of protein into the medium

pEXPR-IBA vectors with Strep-tag® for mammalian expression

pEXPR-IBA vectors from the Classic Cloning series for expression of Strep-tagged proteins in mammalian cells are available with different protease cleavage sites.
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.

 

pEXPR-IBA Features Benefits
Strep-tag®II Purification of recombinant protein using Strep-Tactin® matrices
CMV immediate-early promoter/enhancer High-level expression in a wide range of mammalian cells
Multiple cloning site Insertion of gene of interest and fusion to Strep-tag®. Compatible with pASK-IBA vectors
Neomycin resistance gene Selection of stable transfectants in mammalian cells
Ampicillin resistance gene Selection in E. coli
pUC origin High-copy number replication in E. coli
Factor Xa, thrombin (thb),
enterokinase (ent) and
TEV cleavage sites
Removal of the Strep-tag® if required (generally not necessary)
Yeast

pYSG-IBA vectors

 

  • High-level expression in yeast
  • Tightly regulated expression due to Cu++-inducible CUP1 promoter
  • Ampicillin resistance gene and markers leu2-d and URA3
  • High copy number
Insect cells

pLSG-IBA vectors

 

  • Cloning and expression of recombinant proteins in insect cells
  • Polyhedrin promoter drives high-level expression of the insert
  • pUC ori and ampicillin resistance gene support propagation in E. coli
Dictyostelium disoideum
StarGate Acceptor Vector adaptation StarGate Acceptor Vector adaptation pKOSG-IBA-dicty1 
For convenient and efficient generation of gene knock out vectors for Dictyostelium discoideum.
 
Reference: