VectorBuilder offers many popular vector components that users can choose from when designing their vectors. The tables below provide detailed information on these popular components, which are listed separately by category.
Epitope Tags
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| FLAG | FLAG epitope tag | Facilitates antibody binding of tagged protein. | J Biochem Biophys Methods. 49:455 (2001); Appl Microbiol Biotechnol. 60:523 (2003) | View |
| 3xFLAG | 3 tandem FLAG epitope tag | Facilitates antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| HA | Hemagglutinin epitope tag | Facilitates antibody binding of tagged protein. | Nature. 443:230 (2006) | View |
| Myc | c-Myc epitope tag | Facilitates antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| V5 | V5 epitope tag | Facilitates antibody binding of tagged protein. | J Biol Chem. 282:20799 (2007) | View |
| OVA(257-264) | H-2Kb-restricted OVA MHC class I epitope | Allows detection of a strong CD8+ cytolytic T cell response. | Eur J Immunol. 21:2891 (1991); J Immunol. 150:1212 (1993) | View |
| OVA(323-339) | I-Ad-restricted OVA MHC class II epitope | Allows studying the nature of class II MHC-peptide binding and T-cell activation. | Biochemistry. 38:16663 (1999); J Immunol. 171:1999 (2003) | View |
| 10xGCN4_v4 | The v4 version of the GCN4 peptide array with 10 copies of the peptide binding site, also know as SunTag10x_v4 | Allows tagged proteins to be recognized by the GCN4 scFv antibody. | Cell. 159: 635 (2014) | View |
| Rho1D4 | A 9 amino acid 1D4 epitope derived from the C-terminus of bovine rhodopsin | Allows tagged proteins to be recognized by the Rho1D4 antibody; typically recommended for the purification and characterization of membrane proteins. | Methods Mol Biol. 1177:1 (2014) | View |
| ALFA-tag | ALFA-tag | Allows tagged proteins to be recognized by the NbALFA nanobody. | Nature Communications. 10:4403 (2019) | View |
Purification Tags
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| 6xHis | 6 tandem histidine tag | Facilitates purification by nickel or cobalt matrix and antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| 10xHis | 10 tandem histidine tag | Facilitates purification by nickel or cobalt matrix and antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| Strep-tag_II | Strep-tag II | Facilitates purification by Strep-Tactin (modified streptavidin) matrix and antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| Twin-Strep-tag | 2 tandem Strep-tag II linked with 3xGGGS | Facilitates purification by Strep-Tactin (modified streptavidin) matrix and antibody binding of tagged protein. | Protein Expr Purif. 92:54 (2013) | View |
| S-tag | S-tag | Facilitates purification by S-protein matrix and antibody binding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| MBP | Maltose-binding protein | Facilitates purification by amylose matrix and antibody binding of tagged protein; increases the solubility of tagged protein in bacteria. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| GST | Glutathione S-transferase | Facilitates purification by glutathione matrix and antibody binding of tagged protein; helps to stabilize tagged protein and protect it against intracellular protease cleavage. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| Avi | Avi tag | Facilitates biotinylation of tagged protein by the enzyme BirA so the protein can be isolated by streptavidin. | Protein Sci. 8:921 (1999); Methods Mol Biol. 1266: 171 (2015) | View |
| Halo | HaloTag7 | Increases expression and solubility of tagged protein; enables covalent, efficient and specific capture of a protein of interest onto HaloLinkâ„¢ Resin; high yield and purity. | Protein Expr Purif. 68:110 (2009) | View |
| Halo-TEV | HaloTag7 linked with TEV site | Increases expression and solubility of tagged protein; enables covalent, efficient and specific capture of a protein of interest onto HaloLinkâ„¢ Resin; high yield and purity; can be removed by TEV protease; N-terminal tag. | Protein Expr Purif. 68:110 (2009) | View |
| SUMO | Small ubiquitin-like modifier tag | Facilitates purification by enhancing stability and solubility of tagged proteins; can be specifically recognized by SUMO proteases. | Protein Sci.22(11):1466-77 (2013) | View |
| hIgG-Fc | The constant region (CH2 and CH3 domains) of human immunoglobulin heavy chain | Increases protein yield; allows rapid and simple detection of protein expression; enables simple protein G/A affinity chromatography based purification of Fc-tagged proteins. | EMBO Mol Med. 4:1015 (2012); J Biomed Res. 30: 209 (2016) | View |
| mlgG-Fc | The constant region (CH2 and CH3 domains) of mouse immunoglobulin heavy chain | Increases protein yield; allows rapid and simple detection of protein expression; enables simple protein G affinity chromatography based purification of Fc-tagged proteins. | EMBO Mol Med. 4:1015 (2012); Sci Rep. 6:30171 (2016) | View |
| NpuC(HN) | A modified C-terminal segment of the Nostoc punctiforme (Npu) DnaE split intein | Facilitates purification of N-terminal tagged proteins by intein-mediated cleavage and subsequent release of tagless target proteins induced by a simple pH shift (8.5 to 6.2) which leads to a cyclization reaction of the C-terminal asparagine to form a succinimide ring. | Curr Protoc Protein Sci. 91:5.29.1 (2018) | View |
Protease Cleavage Sites
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| EK | Bovine enterokinase cleavage site | Cleaved by enterokinase, which cuts after lysine in the recognition sequence DDDDK. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| TEV | Tobacco etch virus protease cleavage site | Cleaved by tobacco etch virus protease, which cuts after glutamine in the recognition sequence EXXYXQ(G/S) where X can be any amino acid. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| Thrombin | Thrombin cleavage site | Cleaved by thrombin, which cuts after the fourth residue in the recognition sequence XXP(R/K)XX where XX on the N end are hydrophobic residues and XX on the C end are non-acidic residues. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| Factor-Xa | Factor Xa cleavage site | Cleaved by factor Xa, which cuts after arginine in the recognition sequence I(E/D)GR. | Appl Microbiol Biotechnol. 60:523 (2003) | View |
| HRV3C | Human rhinovirus 3C protease site | Cleaved by human rhinovirus 3C protease, the cleavage site is between Gln and Gly in the recognition sequences LEVLFQ↓GP. | Cell.168(3):427-441 (2017) | View |
Signal Peptides (for directing localization of proteins)
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| SV40-NLS | Simian virus 40 nuclear localization signal | Directs nuclear localization of tagged protein. | Cell. 39:499 (1984) | View |
| 3xNLS | 3 tandem simian virus 40 nuclear localization signal | Directs nuclear localization of tagged protein. | J Immunol. 182: 3173 (2009) | View |
| cMYC-NLS | Nuclear localization signal of human c-MYC gene | Directs nuclear localization of tagged protein. | Mol Cell Biol. 8:4048 (1988); Bioconjug Chem. 26: 1004 (2015) | View |
| BPSV40-NLS | A classical nuclear localization signal (NLS) consists of two clusters of basic residues separated by 10–12 residues (bipartite), NLS-containing cargoes are imported by a heterodimeric import receptor complex composed of importin α and importin β | Directs nuclear localization of tagged protein. | J Biol Chem. 276:1317 (2001); Biophys J. 96:3840 (2009); Nature. 540:144 (2016); Nat Methods. 16:887-893 (2019) | View |
| MITO | Mitochondrial targeting sequence from human cytochrome c oxidase subunit VIII gene | Directs tagged protein to the intermembrane space of mitochondria. | Nature. 358:325 (1992); Proc Natl Acad Sci U S A. 96: 11752 (1999) | View |
| Igκ-sig | Murine Ig kappa chain leader sequence | Directs tagged protein to the secretory pathway. | J Immunol Methods. 152:89 (1992) | View |
| α-Factor | Yeast pheromone α-factor leader sequence | Directs export of tagged protein in yeast into culture medium. | Gene. 54:175 (1987) | View |
| HBM | Honeybee melittin signal peptide | Allows enhanced secretion of tagged proteins from insect cells into the medium. | Gene. 98:177 (1991) | View |
| PelB | Pectate lyase B leader sequence | Directs tagged protein to the bacterial periplasm, where the sequence is removed by a signal peptidase. | Protein Expr Purif. 54:227 (2007) | View |
| NES | Nuclear export signal | Directs nuclear export of tagged protein. | PLoS One.10:e0137652 (2015) | View |
| Lyn | N-terminal myristoylation and palmitoylation sequence from Lyn, a Src family tyrosine kinase | Anchors the tagged protein to the plasma membrane. | J Neurosci Methods. 93:37 (1999); J Cell Biol. 174:255 (2006) | View |
| SNAP25 | A protein which is synthesized on free ribosomes and recruited to the inner surface of plasma membrane after the palimitoylation of specific cystesine residues | Directs the tagged proteins to the inner surface of plasma membrane. | EMBO J. 16:1575 (1997) | View |
| GPI | Plasma membrane glycosylphosphatidylinositol anchoring signal | Anchors glycoprotein to the plasma membrane; helps increase the mobility of membrane proteins therefore supporting signal transduction and cellular transport; plays an important role in creating antigens on cell surface. | Genesis. 44:202 (2006) | View |
| TMD-ST | Transmembrane domain of rat liver sialyltransferase | Directs the tagged protein to Golgi. | J Biol Chem. 270:5483 (1995) | View |
| PDGFR | Platelet-derived growth factor receptor beta transmembrane domain | Anchors the tagged protein to the extracellular side of plasma membrane for displaying, usually works with secretory signal peptide. | Proc Natl Acad Sci U S A. 85:3435 (1988); J Biol Chem. 285:12268 (2010) | View |
| GLuc | Gaussia luciferase secretory signal peptide | Allows the tagged protein to be secreted from cytosol. | Microb Cell Fact. 14:20 (2015) | View |
| IL2-sig | Signal peptide of Homo sapiens interleukin 2 | Facilitates secretion of proteins. | Protein Expr Purif.118:105-12(2016) | View |
| HA-sig | The cleavable signal sequence of influenza hemagglutinin | Enhances localization of proteins to the endoplasmic reticulum membrane or plasma membrane. | J Biol Chem. 267:21995 (1992); Biochem Biophys Res Commun. 409:544 (2011) | View |
Detection Tags
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| TC | Tetracysteine tag | Facilitates detection of tagged protein by biarsenical compounds such as FlAsH-EDT2 and ReAsH-EDT2, which turn fluorescent upon binding to the tag. | Nat Protoc. 5: 1666 (2010) | View |
| ProS2 | Protein S of Myxococcus xanthus | Facilitates detection of tagged protein by anti-ProS2 antibody; increases the solubility and stability of tagged protein in bacteria. | J Mol Biol. 286:1533 (1999) | View |
| TF | E.coli trigger factor, a ribosome-associated chaperone | Assists protein folding; facilitates detection of tagged protein by anti-trigger factor antibody; increases the solubility of tagged protein. | Biochim Biophys Acta. 1803:650 (2010) | View |
| Lifeact | A 17-aa F-actin-binding sequence derived from Saccharomyces cerevisiae | Allows visualization of F-actin dynamics in cells; can be used as a universal marker for F-actin imaging or as an alternative to phalloidin. | Nat Methods. 5:605 (2008) | View |
| SNAP | A 20 kDa engineered variant of the human DNA repair protein O-6-alkylguanine-DNA alkyltransferase, optimized for mammalian expression | Allows to observe localization and translocation of SNAP-tag fusion proteins; reacts specifically and rapidly with benzylguanine (BG) and benzylchloropyrimidine (CP) derivatives, leading to irreversible covalent labeling of the SNAP-tag with a wide variety of synthetic probes in live cells. CLIP and SNAP fusion proteins can be simultaneously and specifically labeled with different synthetic probes within cells. | Nat Biotechnol. 21:86 (2003); Methods Mol Biol. 1266:107 (2015) | View |
| CLIP | An engineered variant of the human DNA repair protein O-6-alkylguanine-DNA alkyltransferase, optimized for mammalian expression | Allows to observe localization and translocation of CLIP-tag fusion proteins; reacts specifically and rapidly with O2-benzylcytosine (BC) derivatives, leading to irreversible covalent labeling of the CLIP-tag with a wide variety of synthetic probes in live cells. CLIP and SNAP fusion proteins can be simultaneously and specifically labeled with different synthetic probes within cells. | Chem Biol. 15:128 (2008) | View |
| ACP | Acyl carrier protein of E. coli | Allows to observe localization and translocation of ACP-tag fusion cell surface proteins which are specifically labeled with CoA derivatives through post-translational modifications catalyzed by the phosphopantetheinyl transferase AcpS. | J Am Chem Soc. 126:8896 (2004) | View |
| HiBiT | A small 11 amino acid peptide that binds LgBiT with high affinity | Facilitates detection and monitoring of tagged endogenous protein in live cells; capable of producing highly active Nanoluc luciferase through high affinity complementation with LgBiT. | Biochem Biophys Rep. 12: 40 (2017); Chem Biol. 13:467 (2018) | View |
Degron Tags
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| DD | Destabilizing domain, mutant derived from human FKBP12 protein. | Targets the tagged protein for proteosomal degradation; addition of a compound, Shield-1, can stabilize the protein and prevents degradation. | Bioorg Med Chem Lett. 18:5941 (2008); PLoS One. 4: e6529 (2009) | View |
| S4D | Sal-like protein 4 (SALL4) degron (S4D) tag | Allows degradation of S4D-tagged proteins in the presence of immunomodulatory drugs (IMiDs) which interact with cereblon (CRBN) in the cullin E3 ubiquitin ligase complex to induce degradation of neosubstrates. | Commun Biol. 3:515 (2020) | View |
Dimerization Domains
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| FKBP(F36V) | FK506-binding protein 12 with F36V mutation | Promotes dimerization of tagged proteins in the presence of dimerizer ligands, e.g. AP20187. | Chem Rev. 110:3315 (2010); Sci Rep. 4: 6127 (2014) | View |
Cell-Penetrating Peptides
| Name | Description | Application Notes | References | Sequence |
|---|---|---|---|---|
| TAT | Amino acids 48–60 of transactivator of transcription protein (Tat) from human immunodeficiency virus type 1 (HIV-1). | Directs tagged protein to pass through the plasma membrane of mammalian cells. | J Biol Chem. 276:5836 (2001); ACS Nano. 8:1972 (2014); Biomol Concepts. 8:131 (2017) | View |
| R8 | Octaarginine tag (8 tandem argenines). | Directs tagged protein to pass through the plasma membrane of mammalian cells. | J Biol Chem. 276:5836 (2001); ACS Nano. 8:1972 (2014); Biomol Concepts. 8:131 (2017) | View |