Background
The pEXP series of expression vectors, developed by Aleksei Lulla, are for parallel cloning of fusion constructs (in particular) for screening of soluble and folded for of your target proteins.
These new vectors are based on the pOP series (and all the old benefits of these and their ancesters pBAT series), but they are of course improved onn the old, not least in their multiple cloning site which is designed for sequence and ligation independent cloning (SLIC).
A few of the key features we have introduced to the pExp series:
- They have octa-His fusion in all vectors. Usually in the N-terminus, but sometimes in the C-terminus as well
- They have all the fusions parners of pOP vectors (GST, MBP, TrxA, Sumo, GB1) but a big set of new ones such as beta-lactamase, Trigger Factor, IF2 etc
- They have TEV protease sites after the fusions
- They have a StrepTag in the C-terminus for many that have an N-terminal His-tag
- The vectors have BsaI site in the 5′ end of the cloning site which cleaves it so that you can get the optimal TEV site for you insert
- The XhoI in the 3′ end of the polylinker is in frame with C-terminal tag, translating to two serines. Just add a stop codon before that if you do not want a C-terminal tag
- The polylinker has a stop codon just after the TEV site, so you can have nice control for your expression trials without any insert, or as a negative control in downstream applications.
Note on vector maps: The maps are rather basic, listing typically all of the fusion protein sector with DNA and protein sequences, and the unique restriction sites both between different part of the fusion protein and the complete polylinker. You can also get file
When using these vectors, please cite them as Aleksei Lulla et. al, unpublished.
The vectors
pExp-NHis |
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| Contains mini-cistron (PMID: 3534891), N-terminal 8xHis followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| pExp-NHis map (PDF) | pExp-NHis sequence (FASTA) | pExp-NHis fusion sequence (FASTA) |
pExp-CHis |
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| Contains mini-cistron (PMID: 3534891), C-terminal 8xHis. Cloning of GOI between NcoI and XhoI or HindIII. | ||
| pExp-CHis map (PDF) | pExp-CHis sequence (FASTA) | pExp-CHis fusion sequence (FASTA) |
pExp-His-AFV-TEV |
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| Contains N-terminal 8xHis, fusion partner: AFV (Acidianus Filamentous Virus 1 protein ORF-99, UniProtKB: Q70LE8, PMID: 19472363) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 14.3 Da | 122 residues | pI: 5.2064 |
| pExp-His-AFV-TEV map (PDF) | pExp-His-AFV-TEV sequence (FASTA) | pExp-His-AFV-TEV fusion sequence (FASTA) |
pExp-His-Bla-TEV |
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| Contains N-terminal 8xHis, fusion partner: Bla (beta-lactamase from the halophile Chromohalobacter sp. 560, UniProKB: Q76LX5, PMID: 20838790) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 42.2 Da | 390 residues | pI: 4.3428 |
| pExp-His-Bla-TEV map (PDF) | pExp-His-Bla-TEV sequence (FASTA) | pExp-His-Bla-TEV fusion sequence (FASTA) |
pExp-His-DsbA-TEV |
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| Contains N-terminal 8xHis, fusion partner: DsbA (leaderless thiol:disulfide interchange protein DsbA from E. coli, UniProtKB: P0AEG4) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 23.9 Da | 213 residues | pI: 6.5694 |
| pExp-His-DsbA-TEV map (PDF) | pExp-His-DsbA-TEV sequence (FASTA) | pExp-His-DsbA-TEV fusion sequence (FASTA) |
pExp-His-DsbA-SS-TEV |
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| Contains N-terminal 8xHis, fusion partner: DsbA (leaderless thiol:disulfide interchange protein DsbA from E. coli, active site cysteines are mutated to serines. | ||
| pExp-His-DsbA-SS-TEV map (PDF) | pExp-His-DsbA-SS-TEV sequence (FASTA) | pExp-His-DsbA-SS-TEV fusion sequence (FASTA) |
pExp-His-DsbC-TEV |
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| Contains N-terminal 8xHis, fusion partner: DsbC (leaderless thiol:disulfide interchange protein DsbC from E. coli, UniProtKB: P0AEG6, PMID: 23607455) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 26.2 Da | 240 residues | pI: 6.7324 |
| pExp-His-DsbC-TEV map (PDF) | pExp-His-DsbC-TEV sequence (FASTA) | pExp-His-DsbC-TEV fusion sequence (FASTA) |
pExp-His-GB1-TEV |
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| Contains N-terminal 8xHis, fusion partner: GB1 (domain B1 of Immunoglobulin G-binding protein G from Streptococcus sp., UniProtKB: P19909, PMID: 9385638) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 8.8 Da | 79 residues | pI: 6.2792 |
| pExp-His-GB1-TEV map (PDF) | pExp-His-GB1-TEV sequence (FASTA) | pExp-His-GB1-TEV fusion sequence (FASTA) |
pExp-His-GST-TEV |
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| Contains N-terminal 8xHis, fusion partner: MBP (Glutathione S-transferase from Schistosoma japonicum, UniProtKB: P08515, PMID: 3047011) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 28.1 Da | 241 residues | pI: 6.8621 |
| pExp-His-GST-TEV map (PDF) | pExp-His-GST-TEV sequence (FASTA) | pExp-His-GST-TEV fusion sequence (FASTA) |
pExp-His-IF2-TEV |
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| Contains N-terminal 8xHis, fusion partner: IF2 (domain I of translation initiation factor IF-2 from E. coli, UniProtKB: P0A705, PMID: 14965771) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 20.4 Da | 181 residues | pI: 9.4958 |
| pExp-His-IF2-TEV map (PDF) | pExp-His-IF2-TEV sequence (FASTA) | pExp-His-IF2-TEV fusion sequence (FASTA) |
pExp-His-MBP-TEV |
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| Contains N-terminal 8xHis, fusion partner: MBP (maltose-binding protein (malE) from E. coli, UniProtKB: P0AEX9, PMID: 10452611) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 43.1 Da | 391 residues | pI: 5.4478 |
| pExp-His-MBP-TEV map (PDF) | pExp-His-MBP-TEV sequence (FASTA) | pExp-His-MBP-TEV fusion sequence (FASTA) |
pExp-His-mCherry-TEV |
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| Contains N-terminal 8xHis, fusion partner: mCherry (mCherry fluorescent protein, UniProtKB: X5DSL3) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 29.4 Da | 259 residues | pI: 6.5046 |
| pExp-His-mCherry-TEV map (PDF) | pExp-His-mCherry-TEV sequence (FASTA) | pExp-His-mCherry-TEV fusion sequence (FASTA) |
pExp-His-MsyB-TEV |
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| Contains N-terminal 8xHis, fusion partner: MsyB (acidic protein MsyB from E. coli, UniProtKB: P25738, PMID: 17374413) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 16.9 Da | 147 residues | pI: 3.7917 |
| pExp-His-MsyB-TEV map (PDF) | pExp-His-MsyB-TEV sequence (FASTA) | pExp-His-MsyB-TEV fusion sequence (FASTA) |
pExp-His-NusA-TEV |
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| Contains N-terminal 8xHis, fusion partner: NusA (N utilization substance protein A (NusA) from E. coli, UniProtKB: P0AFF6, PMID: 10506413) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 57.5 Da | 518 residues | pI: 4.4033 |
| pExp-His-NusA-TEV map (PDF) | pExp-His-NusA-TEV sequence (FASTA) | pExp-His-NusA-TEV fusion sequence (FASTA) |
pExp-His-PpiB-TEV |
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| Contains N-terminal 8xHis, fusion partner: PpiB (peptidyl-prolyl cis-trans isomerase B from E. coli, UniProtKB: P23869, PMID: 19459971 ) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 20.8 Da | 187 residues | pI: 6.5584 |
| pExp-His-PpiB-TEV map (PDF) | pExp-His-PpiB-TEV sequence (FASTA) | pExp-His-PpiB-TEV fusion sequence (FASTA) |
pExp-His-sfGFP-TEV |
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| Contains N-terminal 8xHis, fusion partner: sfGFP (superfolder green fluorescent protein, PMID: 16369541) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 29.5 Da | 262 residues | pI: 6.6866 |
| pExp-His-sfGFP-TEV map (PDF) | pExp-His-sfGFP-TEV sequence (FASTA) | pExp-His-sfGFP-TEV fusion sequence (FASTA) |
pExp-His-Sumo-TEV |
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| Contains N-terminal 8xHis, fusion partner: Sumo (Smt3 from Saccharomyces cerevisiae, UniProtKB: Q12306, PMID: 16084395) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 13.8 Da | 119 residues | pI: 6.1568 |
| pExp-His-Sumo-TEV map (PDF) | pExp-His-Sumo-TEV sequence (FASTA) | pExp-His-Sumo-TEV fusion sequence (FASTA) |
pExp-His-TF-TEV |
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| Contains N-terminal 8xHis, fusion partner: TF (Trigger factor (tig) from E. coli, UniProtKB: P0A850, PMID: 18476832) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 50.8 Da | 455 residues | pI: 4.7485 |
| pExp-His-TF-TEV map (PDF) | pExp-His-TF-TEV sequence (FASTA) | pExp-His-TF-TEV fusion sequence (FASTA) |
pExp-His-TFSumo-TEV |
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| Contains N-terminal 8xHis, fusion partner: TF-Sumo (Trigger factor from E. coli (UniProtKB: P0A850) linked to Sumo (Smt3 from Saccharomyces cerevisiae, UniProtKB: Q12306), PMID: 26297996) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 61.9 Da | 552 residues | pI: 4.7277 |
| pExp-His-TFSumo-TEV map (PDF) | pExp-His-TFSumo-TEV sequence (FASTA) | pExp-His-TFSumo-TEV fusion sequence (FASTA) |
pExp-His-Trx-TEV |
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| Contains N-terminal 8xHis, fusion partner: Trx (thioredoxin (trxA) from E. coli, UniProtKB: P0AA25, PMID: 7763371) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 14.4 Da | 132 residues | pI: 6.2083 |
| pExp-His-Trx-TEV map (PDF) | pExp-His-Trx-TEV sequence (FASTA) | pExp-His-Trx-TEV fusion sequence (FASTA) |
pExp-His-Tsf-TEV |
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| Contains N-terminal 8xHis, fusion partner: Tsf (elongation factor Ts from E. coli, UniProtKB: P0A6P1, PMID: 17608803) followed by TEV protease cleavage site, optional C-terminal StrepII-tag. Cloning of GOI between BsaI and XhoI or HindIII. | ||
| Fusion MW: 33.1 Da | 306 residues | pI: 5.9362 |
| pExp-His-Tsf-TEV map (PDF) | pExp-His-Tsf-TEV sequence (FASTA) | pExp-His-Tsf-TEV fusion sequence (FASTA) |