pOPIN Vector Suite

In-Fusion™ cloning and the pOPIN Vector Suite

The OPPF has produced a versatile suite of expression vectors that, in combination with the In-Fusion™ enzyme, has produced a novel HTP cloning process with the following characteristics:

  • The process is single-step - the PCR product is cloned directly into the final expression vector.
  • The majority of the vectors allow expression of proteins from multiple hosts, i.e. a single vector capable of expression in E. coli, mammalian cell lines (e.g. HEK293T cells) and insect cell lines (e.g. Sf9 cells). This enables parallel screening of constructs in multiple hosts.
  • The process is simple to operate in HTP mode - the In-Fusion™ method has now been used to clone in excess of 1000 PCR products into pOPIN expression vectors with very high efficiency.
  • The 'fusion' site between tag and protein of interest is very precise so the final protein translates bear no unwanted vector, or cloning-derived amino acids.
  • All of the pOPIN vectors contain His-tags which are (with the exception of that from pOPINI) cleavable from the protein of interest, either with 3C protease, SUMO protease or Carboxypeptidase A, for improved crystallization.

The sequences of the pOPIN vectors may be downloaded using the links below (in either FASTA or GenBank format). Right-click on the FASTA (Fasta) or GenBank (Genbank) icons and select "Save As...", or, if you have VectorNTI installed, left-click on the GenBank icon to open in VectorNTI directly. Alternatively, try ApE - A Plasmid Editor from M. Wayne Davis at the Department of Biology, University of Utah. Please contact us if you have trouble using these file formats.

Vector Product E. coli HEK293 Baculo
FastaGenbank pOPINFHIS6-3C-POIYYY
FastaGenbank pOPINEPOI-KHIS6YYY
FastaGenbank pOPINS3CHIS6-SUMO-3C-POIYYY
FastaGenbank pOPINMHIS6-MBP-3C-POIYYY
FastaGenbank pOPINTRXHIS6-TRX-3C-POIYYY
FastaGenbank pOPINJHIS6-GST-3C-POIYYY
FastaGenbank pOPINTFHIS6-TF-3C-POIYYY
FastaGenbank pOPINHALO7HIS6-HALO7-3C-POIYYY
FastaGenbank pOPINMSYBHIS6-MSYB-3C-POIYYY
FastaGenbank pOPINNUSAHIS6-NUSA-3C-POIYYY
FastaGenbank pOPINGSS[RPTPmu]-POI-KHIS6NYY
FastaGenbank pOPINHSS[RPTPmu]-HIS6-3C-POINYY
FastaGenbank pOPINGSSS[RPTPmu]-POI-StrepII-KHIS6NYY
FastaGenbank pOPINTTGneoSS[RPTPmu]-POI-KHIS6NYN
FastaGenbank pOPINTTGneo-3C-eGFPSS[RPTPmu]-POI-3C-eGFP-KHIS6NYN
FastaGenbank pOPINTTGneo-3C-HALO7SS[RPTPmu]-POI-3C-HALO7-KHIS6NYN
FastaGenbank pOPINTTGneo-3C-CD4SS[RPTPmu]-POI-3C-CD4-KHIS6NYN
FastaGenbank pOPINTTGneo-3C-FcSS[RPTPmu]-POI-3C-Fc-KHIS6NYN
FastaGenbank pOPINTTEPOI-KHIS6NYN
FastaGenbank pOPINTTFHIS6-3C-POINYN
FastaGenbank pOPINTTIHIS6-POINYN
FastaGenbank pOPINFSHIS6-3C-POI-StrepIIYYY
FastaGenbank pOPINJBHIS6-GST-3C-POI-BAPYYY
FastaGenbank pOPINFBHIS6-3C-POI-BAPYYY
FastaGenbank pOPINE-3C-eGFPPOI-3C-eGFP-KHIS6YYY
FastaGenbank pOPINE-3C-HALO7POI-3C-HALO7-KHIS6YYY
FastaGenbank pOPINE-3C-CD4POI-3C-CD4-KHIS6YYY
FastaGenbank pOPINRSFHIS6-3C-POIYNN
FastaGenbank pOPINRSJHIS6-GST-3C-POIYNN
FastaGenbank pOPINRSEPOI-KHIS6YNN
FastaGenbank pOPINSHIS6-SUMO-POIYNN
FastaGenbank pOPINAPOI-KHIS6YNN
FastaGenbank pOPINBHIS6-3C-POIYNN
FastaGenbank pOPINKHIS6-GST-3C-POIYNN
FastaGenbank pOPINCDFHIS6-3C-POIYNN
FastaGenbank pOPINCDJHIS6-GST-3C-POIYNN
FastaGenbank pOPINCDMHIS6-MBP-3C-POIYNN
FastaGenbank pOPINCDEPOI-KHIS6YNN
FastaGenbank pOPINMalESS[MalE]-POI-KHIS6YNN
FastaGenbank pOPINDsbASS[DsbA]-POI-KHIS6YNN
FastaGenbank pOPINTolBSS[TolB]-POI-KHIS6YNN
FastaGenbank pOPINPSS[PelB]-POI-KHIS6YNN
FastaGenbank pOPINOSS[OmpA]-POI-KHIS6YNN
FastaGenbank pOPINVHSS[RPTPmu]-POI-VH-KHIS6NYY
FastaGenbank pOPINVLSS[RPTPmu]-POI-VL-KHIS6NYY

For a more detailed printable description of the vector suite and the latest protocols please download the Cloning and Small Scale Expression standard protocols (pdf format).

 

*In-Fusion is a trademark of Clontech Laboratories Inc.