Part:BBa_K1199046:Design
DhaA31-P2A-HheCW249P 1,2,3-tricholoropropane(TCP)->Glycerol
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 835
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 681
Illegal NgoMIV site found at 807
Illegal AgeI site found at 1600 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
There are restriction enzyme sites BglII and KpnI in the sides of P2A peptide sequence.
Source
DhaA31 is from Rhodococcus sp ,and HheC is from Agrobacterium radiobacter
References
1 Maryna Lahoda, Radka Chaloupkova, Alena Stsiapanava, Jiri Damborsky, and Ivana Kuta Smatanovaa, Crystallization and crystallographic analysis of the Rhodococcus rhodochrous NCIMB 13064 DhaA mutant DhaA31 and its complex with 1,2,3-trichloropropane ,Acta Crystallogr Sect F Struct Biol Cryst Commun, 2011 February 25 .
2 Fabritz S, Maaß F, Avrutina O, Heiseler T, Steinmann B, Kolmar H, A sensitive method for rapid detection of alkyl halides and dehalogenase activity using a multistep enzyme assay. AMB Express. 2012 Sep 24.
3 Jeffrey H. Lutje Spelberg, Johan E. T. van Hylckama Vlieg, Tjibbe Bosma, Richard M. Kellogg and Dick B. Janssen, A tandem enzyme reaction to produce optically active halohydrins, epoxides and diols, aDepartment of Biochemistry, Groningen Biomolecular Sciences & Biotechnology Institute, University of Groningen, bDepartment of Organic & Molecular Inorganic Chemistry, University of Groningen, Nijenborgh 4, 9747 AG, Groningen, 26 July 1999 .
4 Assis HMS, Sallis PJ, Bull AT, Hardman DJ. Biochemical characterization of a haloalcohol dehalogenase from Arthrobacter erithii H10a. Enzyme Microb Technol 1998;22:568–74
5 Lixia Tang, Xuechen Zhu, Huayu Zheng, Rongxiang Jiang, and Maja Majerić Elenkov, Key Residues for Controlling Enantioselectivity of Halohydrin Dehalogenase from Arthrobacter sp. Strain AD2, Revealed by Structure-Guided Directed Evolution, Appl Environ Microbiol. 2012 April
6 Ekaterina Minskaia and Martin D. Ryan, Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues,Biomed Res Int. 2013; 2013: 291730.