Protein Expression and Purification Core Facility

Expression vectors

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Expression vectors for E. coli

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Vectors that are commercially available can only be shared with internal EMBL users, but vectors generated at EMBL are freely available to the entire academic research community via a Material Transfer Agreement. Vectors that can be shared with external users are indicated in yellow in the vector tables.

Name Map Antibiotic resistance Promoter Fusion tags Protease cleavage site Source Remarks
pACE1 Sequence Amp T7/lac None None Imre Berger MultiColi acceptor vector
pACE2 Sequence Tet T7/lac None None Imre Berger MultiColi acceptor vector
pACYC177 Sequence Amp + Kan None None None NEB Cloning vector with p15A ori
pACYC-Duet Sequence Cam T7/lac MCS1: His6, MCS2: S-tag None Novagen Duet vector with P15 ori for co-expression
pASK75 Sequence Amp tet Nt OmpA signal sequence, Ct Strep None Biometra
pBAD-His-A Sequence Amp araBAD Nt His6-T7tag-Xpress Enterokinase Invitrogen
pBAD-His-B Sequence Amp araBAD Nt His6-T7tag-Xpress Enterokinase Invitrogen
pBAD-His-C Sequence Amp araBAD Nt His6-T7tag-Xpress Enterokinase Invitrogen
pBAD-His-MCS Sequence Amp araBAD None None A. Geerlof
pBADM-11 Sequence Amp araBAD Nt His6 TEV A. Geerlof
pBADM-20 Sequence Amp araBAD Nt TrxA-His6 TEV A. Geerlof
pBADM-20(+) Sequence Amp araBAD Nt TrxA-His6 TEV A. Geerlof 2 stop codons in MCS
pBADM-30 Sequence Amp araBAD Nt His6-GST TEV A. Geerlof
pBADM-30(+) Sequence Amp araBAD Nt His6-GST TEV A. Geerlof 2 stop codons in MCS
pBADM-40 (+) Sequence Amp araBAD Nt MBP TEV A. Geerlof 2 stop codons in MCS
pBADM-41 (+) Sequence Amp araBAD Nt His6-MBP TEV A. Geerlof 2 stop codons in MCS
pBADM-52 Sequence Amp araBAD Nt LL-DsbA-His6 TEV A. Geerlof
pBADM-52(+) Sequence Amp araBAD Nt LL-DsbA-His6 TEV A. Geerlof
pBADM-60 Sequence Amp araBAD Nt NusA-His6 TEV A. Geerlof
pBADM-60(+) Sequence Amp araBAD Nt NusA-His6 TEV A. Geerlof 2 stop codons in MCS
pBAT4 Sequence Amp T7/lac None None M Hyvonen
pBAT5 Sequence Amp T7/lac None None M Hyvonen
pCal-n Sequence Amp T7/lac Nt CBP Thrombin Stratagene
pCDF-1b Sequence Spec/ Strep T7/lac Nt His6, S-tag Enterokinase Novagen
pCDF-Duet Sequence Spec/ Strep T7/lac MCS1: His6, MCS2: S-tag None Novagen Duet vector with CloDF13 (CDF) ori for co-expression
pDC Sequence Cam T7/lac None None Imre Berger MultiColi donor vector
pDEST 14 Sequence Amp + Cam T7 None None Invitrogen Gateway destination vector
pDEST 15 Sequence Amp + Cam T7 Nt GST None Invitrogen Gateway destination vector
pDEST 17 Sequence Amp + Cam T7 Nt His6 None Invitrogen Gateway destination vector
pDK Sequence Kan lac None None Imre Berger MultiColi donor vector
pDNR-Dual Sequence Amp + Cam T7 Nt (His-Asn)6 None Clontech Donor vector
pDONR201 Sequence Kan + Cam None None None Invitrogen Gateway donor vector
pDONR207 Sequence Gent + Cam None None None Invitrogen Gateway donor vector
pDS Sequence Spect lac None None Imre Berger MultiColi donor vector
pENTR4 Sequence Kan + Cam None None None Invitrogen Gateway entry (Dual Selection) vector
pET-3a Sequence Amp T7 Nt T7-tag None Novagen
pET-3b Sequence Amp T7 Nt T7-tag None Novagen
pET-3c Sequence Amp T7 Nt T7-tag None Novagen
pET-3d Sequence Amp T7 Nt T7-tag None Novagen
pET-12a Sequence Amp T7 Nt OmpT signal sequence None Novagen For expression of secreted proteins in E. coli
pET-14b Sequence Amp T7 Nt His6 Thrombin Novagen
pET-15b Sequence Amp T7/lac Nt His6 Thrombin Novagen
pET-16b Sequence Amp T7/lac Nt His10 Factor Xa Novagen
pET-19b Sequence Amp T7/lac Nt His10 Enterokinase Novagen
pET-20b(+) Sequence Amp T7 Nt pelB signal sequence, Ct His None Novagen For expression of secreted proteins in E. coli
pET-21d(+) Sequence Amp T7/lac Nt T7-tag, Ct His6 Novagen
pET-22b(+) Sequence Amp T7/lac Nt pelB signal sequence, Ct His None Novagen For expression of secreted proteins in E. coli
pET-24(+) Sequence Kan T7/lac Ct His6 None Novagen For expression of genes with bacterial translation signals (lacks RBS and ATG start codon)
pET-24a(+) Sequence Kan T7/lac Nt T7-tag, Ct His6 None Novagen Sometimes also referred to as pET24a
pET-24d(+) Sequence Kan T7/lac Nt T7-tag, Ct His6 None Novagen Sometimes also referred to as pET24d
pET-28a(+) Sequence Kan T7/lac Nt His6-thrombin-T7tag, Ct His6 Thrombin Novagen
pET-28c(+) Sequence Kan T7/lac Nt His6-thrombin-T7tag, Ct His6 Thrombin Novagen
pET-32a(+) Sequence Amp T7/lac Nt TrxA-His6-thrombin-S-enterokinase, Ct His6 Thrombin, enterokinase Novagen
pET-32b(+) Sequence Amp T7/lac Nt TrxA-His6-thrombin-S-enterokinase, Ct His6 Thrombin, enterokinase Novagen
pET-32c(+) Sequence Amp T7/lac Nt TrxA-His6-thrombin-S-enterokinase, Ct His6 Thrombin, enterokinase Novagen
pET-39b(+) Sequence Kan T7/lac Nt DsbA-His6-thrombin-S-enterokinase, Ct His8 Thrombin, enterokinase Novagen
pET-40b(+) Sequence Kan T7/lac Nt DsbC-His6-thrombin-S-enterokinase, Ct His8 Thrombin, enterokinase Novagen
pET28-Sumo1eGFP Sequence Kan T7/lac Nt His6-thrombin-hSumo1 Thrombin, hSenP2 Hüseyin Besir
pETcoco-1 Sequence Cam T7/lac Nt His6-S-tag, Ct HSV-tag, Ct His6 Enterokinase Novagen Allows "on command" amplification of vector copy number from single to medium copy
pETDsbA_1b Sequence Kan T7/lac Nt DsbA-His6-TEV-eYFP TEV G. Stier
pETDsbAin_1b Sequence Kan T7/lac Nt LL_DsbA-His6-TEV-eYFP TEV G. Stier
pET-Duet Sequence Amp T7/lac MCS1: His6, MCS2: S-tag None Novagen Duet vector with ColE1 ori for co-expression
pETG-10A Sequence Amp + Cam T7/lac Nt His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-10K Sequence Kan + Cam T7/lac Nt His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-20A Sequence Amp + Cam T7/lac Nt TrxA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-20K Sequence Kan + Cam T7/lac Nt TrxA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-30A Sequence Amp + Cam T7/lac Nt His6-GST, Ct His6 None A. Geerlof Gateway destination vector
pETG-30K Sequence Kan + Cam T7/lac Nt His6-GST, Ct His6 None A. Geerlof Gateway destination vector
pETG-40A Sequence Amp + Cam T7/lac Nt MBP, Ct His6 None A. Geerlof Gateway destination vector
pETG-40K Sequence Kan + Cam T7/lac Nt MBP, Ct His6 None A. Geerlof Gateway destination vector
pETG-41A Sequence Amp + Cam T7/lac Nt His6-MBP, Ct His6 None A. Geerlof Gateway destination vector
pETG-41K Sequence Kan + Cam T7/lac Nt His6-MBP, Ct His6 None A. Geerlof Gateway destination vector
pETG-50A Sequence Amp + Cam T7/lac Nt DsbA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-50K Sequence Kan + Cam T7/lac Nt DsbA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-52A Sequence Amp + Cam T7/lac Nt LL-DsbA-His6 None A. Geerlof Gateway destination vector
pETG-52K Sequence Kan + Cam T7/lac Nt LL-DsbA-His6 None A. Geerlof Gateway destination vector
pETG-60A Sequence Amp + Cam T7/lac Nt NusA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETG-60K Sequence Kan + Cam T7/lac Nt NusA-His6, Ct His6 None A. Geerlof Gateway destination vector
pETGB1_1a Sequence Kan T7/lac Nt His-GB1-TEV-eYFP, Ct His6 TEV G. Stier
pETGB1_1c Sequence Kan T7/lac Nt GB1-TEV-eYFP, Ct His6 TEV G. Stier
pETGB1_dirMa Sequence Kan T7/lac Nt GB1-eYFP, Ct His6 None G. Stier
pETGST_1a Sequence Kan T7/lac Nt His6-GST-TEV-eYFP, Ct His6 TEV G. Stier
pETGST_1b Sequence Kan T7/lac Nt GST-His6-TEV-eYFP, Ct His6 TEV G. Stier
pETGST_dirMa Sequence Kan T7/lac Nt His6-GST-eYFP, Ct His6 None G. Stier
pETHis_1a Sequence Kan T7/lac Nt His6-TEV-eYFP, Ct His6 TEV G. Stier
pETM10 Sequence Kan T7/lac Nt His6, Ct His6 None G. Stier contains EF-hands 3,4 from alpha-actinin insert in modified pET-24d
pETM11 Sequence Kan T7/lac Nt His6, Ct His6 TEV G. Stier contains MXD1 insert in modified pET-24d
pETM11-SUMO3eGFP Sequence Kan T7/lac Nt His6-TEV-hSUMO3 TEV, hSENP2 H. Besir pETM11 with E. coli codon optimized synthetic sumo3 gene
pETM12 Sequence Kan T7 Nt His6 Thrombin G. Stier contains Actin-binding domain insert in modified pET-9d
pETM13 Sequence Kan T7/lac Ct His6 None G. Stier contains Actin-binding domain insert in modified pET-24d
pETM14 Sequence Kan T7/lac Nt His6, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM20 Sequence Amp T7/lac Nt TrxA- His6, Ct His6 TEV G. Stier contains hAGER (RAGE) insert in modified pET-32
pETM22 Sequence Kan T7/lac Nt TrxA- His6, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM30 Sequence Kan T7/lac Nt His6-GST, Ct His6 TEV G. Stier contains PCBD1 (PHS) insert in modified pET-24d
pETM33 Sequence Kan T7/lac Nt His6-GST, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM40 Sequence Kan T7/lac Nt MBP, Ct His6 TEV G. Stier modified pET-24d; contains two STOP codons in MCS
pETM41 Sequence Kan T7 Nt His6-MBP, Ct His6 TEV G. Stier modified pET-9d; contains two STOP codons in MCS; insertions in backbone?
pETM43 Sequence Kan T7/lac Nt MBP, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM44 Sequence Kan T7/lac Nt His6-MBP, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM50 Sequence Kan T7/lac Nt DsbA-His6, Ct His6 TEV G. Stier contains hAGER (RAGE) insert in modified pET-24d; leader sequence for export to periplasm
pETM51 Sequence Kan T7/lac Nt DsbA-His6, Ct His6 Thrombin G. Stier contains hAGER (RAGE) insert in modified pET-24d; leader sequence for export to periplasm
pETM52 Sequence Kan T7/lac Nt LL-DsbA-His6, Ct His6 TEV G. Stier contains hAGER (RAGE) insert in modified pET-24d; leaderless (LL) DsbA fusion for cytoplasmic expression
pETM54 Sequence Kan T7/lac Nt DsbA-His6, Ct His6 3C A. Geerlof modified pET-24d; leader sequence for export to periplasm; contains two STOP codons in MCS
pETM55 Sequence Kan T7/lac Nt LL DsbA-His6, Ct His6 3C A. Geerlof modified pET-24d; leaderless (LL) DsbA fusion for cytoplasmic expression; contains two STOP codons in MCS
pETM60 Sequence Kan T7/lac Nt NusA-His6, Ct His6 TEV G. Stier contains Actin-binding domain insert in modified pET-24d
pETM66 Sequence Kan T7/lac Nt NusA-His6, Ct His6 3C A. Geerlof modified pET-24d; contains two STOP codons in MCS
pETM70 Sequence Kan T7/lac Nt CBP, Ct His6 TEV A. Geerlof modified pET-24d
pETM80 Sequence Kan T7/lac Nt DsbC-His6 TEV G. Stier modified pET-24d; leader sequence for export to periplasm
pETM82 Sequence Kan T7 Nt LL-DsbC-His6 TEV G. Stier modified pET-9d; leaderless (LL) DsbC fusion for cytoplasmic expression
pETMBP_1a Sequence Kan T7/lac Nt His6-MBP-TEV-eYFP, Ct His6 TEV G. Stier
pETMBP_1b Sequence Kan T7/lac Nt MBP-His6-TEV-eYFP, Ct His6 TEV G. Stier
pETNusA_1a Sequence Kan T7/lac Nt His6-NusA-TEV-eYFP, Ct His6 TEV G. Stier
pETNusA_dirMa Sequence Kan T7/lac Nt His6-NusA-eYFP, Ct His6 None G. Stier
pETtrx_1a Sequence Kan T7/lac Nt His6-TrxA-TEV-eYFP, Ct His6 TEV G. Stier
pETtrx_1b Sequence Kan T7/lac Nt TrxA-His6-TEV-eYFP, Ct His6 TEV G. Stier
pETZ1_1b Sequence Kan T7/lac Nt Ztag-His6-TEV-eYFP, Ct His6 TEV G. Stier The Z-tag is based on the Staphylococcus protein A and can bind to the Fc-fragment of antibodies
pETZ2_1a Sequence Kan T7/lac Nt His6-Ztag-TEV-eYFP, Ct His6 TEV G. Stier The Z-tag is based on the Staphylococcus protein A and can bind to the Fc-fragment of antibodies
pETZ2_1b Sequence Kan T7/lac Nt Ztag-His6-TEV-eYFP, Ct His6 TEV G. Stier The Z-tag is based on the Staphylococcus protein A and can bind to the Fc-fragment of antibodies
pETZt2_1a Sequence Kan T7/lac Nt Z2tag-His6-TEV-eYFP, Ct His6 TEV G. Stier The Z-tag is based on the Staphylococcus protein A and can bind to the Fc-fragment of antibodies
pGAT Sequence Amp T7/lac Nt His6-GST None M. Hyvonen
pGAT2 Sequence Amp T7/lac Nt His6-GST Thrombin M. Hyvonen
pGEX-3X Sequence Amp tac/lac Nt GST Factor Xa Pharmacia
pGEX-4T-1 Sequence Amp tac/lac Nt GST Thrombin Pharmacia
pGEX-4T-2 Sequence Amp tac/lac Nt GST Thrombin Pharmacia
pGEX-4T-3 Sequence Amp tac/lac Nt GST Thrombin Pharmacia
pGEX-5X-1 Sequence Amp tac/lac Nt GST Factor Xa Pharmacia
pGEX-5X-2 Sequence Amp tac/lac Nt GST Factor Xa Pharmacia
pGEX-5X-3 Sequence Amp tac/lac Nt GST Factor Xa Pharmacia
pGEX-6P-1 Sequence Amp tac/lac Nt GST 3C Pharmacia
pGEX-6P-2 Sequence Amp tac/lac Nt GST 3C Pharmacia
pGEX-6P-3 Sequence Amp tac/lac Nt GST 3C Pharmacia
pHAT Sequence Amp T7/lac Nt His6 None M Hyvonen
pHAT2 Sequence Amp T7/lac Nt His6 None M Hyvonen
pKK223-3 Sequence Amp tac/lac None None Pharmacia
pKK233-2 Sequence Amp trc/lac None None Pharmacia
pLysS Sequence Cam tet, T7 φ3.8 None None Merck The T7 lysozyme coding sequence is in the antisense orientation relative to the tet promoter, so only a small amount of T7 lysozyme is produced.
pMal-c2 Sequence Amp tac/lac Nt MBP Factor Xa NEB no signal sequence
pMal-p2 Sequence Amp tac/lac Nt malE signal sequence, Nt MBP Factor Xa NEB malE signal sequence
pPCR-Script Amp SK(+) Sequence Amp T7, T3 None None Stratagene Derivative of pBluescript II SK(+) with an SrfI site for inserting blunt PCR products
pPCR-Script eGFP Sequence Amp T7, T3 eGFP inserted in LacZa None A. Geerlof Derivative of pBluescript II SK(+) with an SrfI site for inserting blunt PCR products with eGFP stuffer fragment
pProEx HTa Sequence Amp trc/lac Nt His6 TEV Life Technologies
pProEx HTb Sequence Amp trc/lac Nt His6 TEV Life Technologies
pProEx HTc Sequence Amp trc/lac Nt His6 TEV Life Technologies
pQE-16 Sequence Amp T5/lac Nt DHFR-His6 Qiagen
pQE-30 Sequence Amp T5/lac Nt His6 Qiagen
pQE-31 Sequence Amp T5/lac Nt His6 Qiagen
pQE-32 Sequence Amp T5/lac Nt His6 Qiagen
pQE-60 Sequence Amp T5/lac Ct His6 Qiagen
pQE-70 Sequence Amp T5/lac Ct His6 Qiagen
pQE-80L Sequence Amp T5/lac Nt His6 Qiagen
pQE-81L Sequence Amp T5/lac Nt His6 Qiagen
pQE-82L Sequence Amp T5/lac Nt His6 Qiagen
pREP4 Sequence Kan lacI None None Qiagen Bacterial plasmid with p15A origin and lacI gene for regulating expression from pQE vectors
pRSET A Sequence Amp T7 Nt His6-T7tag-Xpress Enterokinase Thermo
pRSET B Sequence Amp T7 Nt His6-T7tag-Xpress Enterokinase Thermo
pRSET C Sequence Amp T7 Nt His6-T7tag-Xpress Enterokinase Thermo
pRSF-1b Sequence Kan T7/lac Nt His6, Ct S-tag Enterokinase Novagen Bacterial vector with an RSF 1030 origin for the regulated expression of N-terminally 6xHis-tagged proteins
pRSFDuet Sequence Kan T7/lac MCS1: His6, MCS2: S-tag Novagen Duet vector with an RSF 1030 origin for the co-expression of two genes
pSP73 Sequence Amp SP6, T7 RNA polymerase None None Cloning vector for in vitro transcription using the SP6 and T7 RNA polymerase promoters
pTA Plus Sequence Amp T7, T3 None None A. Geerlof TA cloning vector with 3'-T overhangs derived from pPCR-Script Amp
pTrcHis2 A Sequence Amp trc/lac Ct Myc-His6 Thermo Includes a synthetic minicistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream gene
pTrcHis2 B Sequence Amp trc/lac Ct Myc-His6 Thermo Includes a synthetic minicistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream gene
pTrcHis2 C Sequence Amp trc/lac Ct Myc-His6 Thermo Includes a synthetic minicistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream gene
pTrcHis2/LacZ Sequence Amp trc/lac LacZ, Ct Myc-His6 Thermo Includes a synthetic minicistron containing a ribosome binding site (Shine-Dalgarno sequence), for enhancing the bacterial expression of a downstream gene
pZA31-Luc Sequence Cam PLtetO-1 None None H. Bujard Contains luciferase gene
pZE12-Luc Sequence Amp PLlacO-1 None None H. Bujard Contains luciferase gene
pZE21-MCS-1 Sequence Kan PLtetO-1 None None H. Bujard
pZS*24-MCS-1 Sequence Kan Plac/ara-1 None None H. Bujard