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Strep•Tag® II Expression Vectors  | | | | • New Strep•Tag II vectors | | • Vector Table | | • Vector Maps | | Back to Strep•Tag/Strep•Tactin Affinity Purification page | | | | New Strep•Tag II vectors | | | | Novagen latest enhancement of the pET (E. coli), pIEx™ (insect cells) and pTriEx™ (multi-system: E. coli, baculovirus and mammalian) expression vector series involves the incorporation of the proven Strep•Tag II fusion technology (1). The short 8 aa Strep•Tag II coding sequence was added to these vectors as an N-terminal fusion tag just upstream of an enterokinase or HRV 3C cleavage site. These vectors share several additional features: they provide a multiple cloning site (MCS) for traditional cloning, they encode an optional C-terminal 10x His•Tag ® sequence, and all carry the resistance marker to ampicillin. Vectors featuring the HRV 3C protease site also encode an optional C-terminal thrombin protease site for removal of the C-terminal His · Tag sequence. The presence of “gentle elution” tags at both the N-terminus (Strep•Tag II) and C-terminus (optional His · Tag) make these vectors ideal for dual purification strategies designed to isolate full-length, highly purified fusion proteins. | | | | Ligation Independent Cloning (LIC) Options : | | pTriEx-8 are all new members of the Radiance™ Cloning System based on Ek/LIC cloning while pET-52b, pIEx-9 and pTriEx-6 are available in a new 3C/LIC format. The single-stranded overhangs present in the 3C/LIC system complete the HRV 3C protease site coding sequence upstream of the insert and initiate the thrombin protease site coding sequence downstream of the insert. This new configuration allows the removal of the N-terminal and/or C-Terminal fusion tags by the respective protease if desired. | | | | Secretion Options : | | pIEx-10 and pTriEx-7 feature the mouse IgM signal sequence for the secretion of fusion proteins. This signal sequence has previously been demonstrated to function in both insect (2) and mammalian cells (3) and was verified with our constructs using Rluc recombinants. In addition, the signal sequence is also functional in E. coli. | | | | Vector Table | | | Host and Promoter System | N-Terminal Features | C-Terminal Features | Antibiotic Resistance | | Vector | E. coli
T7lac promoter | Insect cells (Transient)
hr5 enhancer, ie1 promoter | Insect cells (Baculovirus)
p10 promoter |
CMV ie enhancer and promoter
| Secretion Signal | | Ek/LIC site | 3C/LIC site | Thrombin site | 10x His•Tag | Ap | | pET-51b(+) | X | | | | | X | X | | | X | X | | pET-52b(+) | X | | | | | X | | X | X | X | X | | pIEx-8 | | X | | | | X | X | | | X | X | | pIEx-9 | | X | | | | X | | X | X | X | X | | pIEx-10 | | X | | | X | X | X | | | X | X | | pTriEx-5 | X | | X | X | | X | X | | | X | X | | pTriEx-6 | X | | X | X | | X | | X | X | X | X | | pTriEx-7 | X | | X | X | X | X | X | | | X | X |
| | | | Vector Maps | | |  | | |  | | |  | | | References 1) IBA (will give you the patent number) Kim, H. G., Yang, S. M., Lee, Y. C., Do, S .I., Chung, I. S., and Yang, J. M. (2003)
2) Biochem. Biophys. Res. Comm. 305, 488-493
3) Kurosawa, N., Hamamoto, T., Lee, Y. C., Nakaoka, T., Kojima, N., Tsuji, S., (1994), J. Biol. Chem. 269, 1402-1409. |
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