Protein domains/Affinity
Protein tags and modifiers are short peptide sequences cloned in frame with protein coding sequences that change the protein's behavior. Affinity tags are generally appended to the N- or C-terminus of proteins to enable ready purification of the protein from cells.
Generally, cells expressing the protein fused to an affinity tag are pelleted, lysed, and applied to a column, resin or other solid support that displays a ligand to the affinity tags. The solid support is washed several times with buffer to eliminate unbound proteins. Then the protein of interest is eluted from the solid support via a buffer that causes the affinity tag to dissociate from the ligand resulting in a purified protein.
There are several different affinity tags available for purification. Each has benefits and advantages depending on your protein of interest and the cell from which you are purifying it.
Head domains
| Name | Description | Column/resin | AA sequence | Length |
|---|---|---|---|---|
| BBa_I712007 | c-Myc affinity tag | 30 | ||
| BBa_I712008 | HA affinity tag | 30 | ||
| BBa_K112703 | His affinity tag | 21 | ||
| BBa_K112705 | S affinity tag | 48 | ||
| BBa_K133035 | Methionine-His affinity tag | 21 | ||
| BBa_T2001 | 7x His tag head domain | 27 | ||
| BBa_T2004 | FLAG octapeptide head domain (DYKDDDDK) | 30 | ||
| BBa_T2007 | c-Myc head domain (EQKLISEEDL) | 36 | ||
| BBa_T2010 | hemagglutinin (HA) tag head domain (YPYDVPDYA) | 33 | ||
| BBa_T2013 | Strep-tag II head domain (WSHPQFEK) | 30 | ||
| BBa_T2016 | S-tag head domain (KETAAAKFERQHMDS) | 51 |
Tail domains
| Name | Description | Column/resin | AA sequence | Length |
|---|---|---|---|---|
| BBa_J64005 | 3x FLAG affinity tag | 81 | ||
| BBa_K112503 | < myc ! in BBb | 33 | ||
| BBa_K112507 | < HA ! in BBb | 33 | ||
| BBa_K112601 | < AP tag ! in BBb | 48 | ||
| BBa_K112603 | < FLAG tag ! in BBb | 27 | ||
| BBa_K1319007 | 6xHis tag with double stop codon | 24 | ||
| BBa_K133036 | RGD-His affinity tag-Stop | 30 | ||
| BBa_K133133 | His affinity tag-stop | 21 | ||
| BBa_K525710 | DNA ligase from Escherichia coli (LigA) with PT7, RBS and His-tag | 2064 | ||
| BBa_T2000 | 7x His tag tail domain | 27 | ||
| BBa_T2003 | FLAG octapeptide tail domain (DYKDDDDK) | 30 | ||
| BBa_T2006 | c-Myc tail domain (EQKLISEEDL) | 36 | ||
| BBa_T2009 | hemagglutinin (HA) tag tail domain (YPYDVPDYA) | 33 | ||
| BBa_T2012 | Strep-tag tail domain (WRHPQFGG) | 30 | ||
| BBa_T2015 | S-tag tail domain (KETAAAKFERQHMDS) | 51 | ||
| BBa_T2021 | VSV-G epitope tag tail domain (YTDIEMNRLGK) | 39 |
Internal domains
| Name | Description | Column/resin | AA sequence | Length |
|---|---|---|---|---|
| BBa_J32017 | S-Tag | 46 | ||
| BBa_K112502 | < myc > in BBb | 30 | ||
| BBa_K112506 | < HA > in BBb | 30 | ||
| BBa_K112602 | < FLAG tag > in BBb | 24 | ||
| BBa_K128000 | FLAG affinity tag | 25 | ||
| BBa_K128001 | HA affinity tag | 27 | ||
| BBa_K128005 | His affinity tag | 18 | ||
| BBa_K3765000 | SpyTag002 | 42 | ||
| BBa_T2002 | 7x His tag special internal domain | 21 | ||
| BBa_T2005 | FLAG octapeptide special internal domain (DYKDDDDK) | 24 | ||
| BBa_T2008 | c-Myc special internal domain (EQKLISEEDL) | 30 | ||
| BBa_T2011 | hemagglutinin (HA) tag special internal domain (YPYDVPDYA) | 27 | ||
| BBa_T2014 | Strep-tag II special internal domain (WSHPQFEK) | 24 | ||
| BBa_T2017 | S-tag special internal domain (KETAAAKFERQHMDS) | 45 | ||
| BBa_T2022 | VSV-G epitope tag special internal domain (YTDIEMNRLGK) | 33 | ||
| BBa_T2025 | Glutathione S-transferase internal domain | 657 | ||
| BBa_T2028 | Chitin binding internal domain | 156 |
Head or tail or internal
| Name | Description | Column/resin | AA sequence | Length |
|---|---|---|---|---|
| BBa_I757013 | His affinity tag | 36 | ||
| BBa_I757014 | Strep-Tag II affinity tag | 24 | ||
| BBa_J18909 | His tag (6xHis; E. coli - optimized) | 18 | ||
| BBa_J18916 | Strep tag II | 24 | ||
| BBa_J18917 | GST tag (E. coli) | 687 | ||
| BBa_J70620 | RFC12 Maltose Binding Protein Internal Domain [malE] | 1140 | ||
| BBa_K1404006 | p70-CsgA, curli generator | 547 | ||
| BBa_K1404007 | p70-CsgA-His, His-tagged curli generator | 577 | ||
| BBa_K1404008 | p70-CsgA-His*2, double His-tagged curli generator | 607 | ||
| BBa_K157011 | His affinity tag; Freiburg standard | 18 | ||
| BBa_K157012 | Strep affinity tag II; Freiburg standard | 24 | ||
| BBa_K2144101 | Vector containing Hisx6 and Sortase tag LPETGG | 2117 | ||
| BBa_K2812000 | Carbohydrate-binding domain from MpIBP | 576 | ||
| BBa_K3204001 | C-LytAm7 Affinity Polypeptide Tag | 411 | ||
| BBa_K3765003 | 10xHis Tag | 30 | ||
| BBa_K404211 | ViralBrick-587KO-His-Tag | 105 | ||
| BBa_K4359007 | V5 Tag | 42 | ||
| BBa_K4378000 | TphB: An effective overexpression casette | 1080 | ||
| BBa_K4422000 | Internal c-Myc (Pichia pastoris codon optimized) | 30 | ||
| BBa_K823034 | 3x FLAG tag (Freiburg standard+RBS) | 66 | ||
| BBa_K823035 | HA-tag (Freiburg standard+RBS) | 27 | ||
| BBa_K823036 | cMyc-tag (Freiburg standard+RBS) | 30 | ||
| BBa_K823037 | 10x His-tag (Freiburg standard+RBS) | 30 | ||
| BBa_K823038 | Strep-tag (Freiburg standard+RBS) | 24 | ||
| BBa_S04282 | K157011(His affinity tag; Freiburg standard):K157004(Fluorescin-A-binding ) | 548 |
References
<biblio>
- Kimple pmid=18429272
- Arnau pmid=16427311
- Azarkan pmid=17113368
- Waugh pmid=15922084
</biblio>