Experience: Works
Not Used
Coding
Part:BBa_K1497032
Designed by: Sascha Hein, Rene Sahm, Malte Blumenroth, Tobias Gabriel, Sebastian Palluk Group: iGEM14_TU_Darmstadt (2014-10-07)
Scaffold (G-S-P-His-Cys)
Usage and Biology
The protein scaffold consists of three different protein binding domains namely the GBD (BBa_K1497024), SH3 (BBa_K1497025) and PDZ (BBa_K1497026) domains. The domains can be linked together in any number and in any order. Therefore, a broad variety of scaffold proteins can be constructed from the initial domains depending on the application. The scaffold protein shown here consists of all domains in the order GBD1SH31PDZ1 or GSP for short. The coding sequence was optimized for the expression in E. coli and revised for the usage as a BioBrick. Additionally, a C-terminal His-tag was added allowing easy purification of the protein.
In this BioBrick, a cysteine residue was added directly behind the His-tag. The thiole group of the cysteine can be used for covalent crosslinking of the scaffold by reactions with maleimids and haloacetamids. The scaffold itself contains no other cysteine residues. Thus, coupling should be regiospecific.
Assembly Compatibility:
- 10
- 12
- 21
INCOMPATIBLE WITH RFC[21] Illegal XhoI site found at 865
- 23
- 25
INCOMPATIBLE WITH RFC[25] Illegal AgeI site found at 76 Illegal AgeI site found at 199
- 1000
COMPATIBLE WITH RFC[1000]
References
Dueber, John E.; Wu, Gabriel C.; Malmirchegini, G. Reza; Moon, Tae Seok; Petzold, Christopher J.; Ullal, Adeeti V. et al. (2009): Synthetic protein scaffolds provide modular control over metabolic flux. In Nat. Biotechnol. 27 (8), pp. 753–759. DOI: 10.1038/nbt.1557.
Parameters
n/a | Scaffold (G-S-P-His-Cys) |
| |