Part:BBa_K4496001:Design
Optimized Jararhagin for Escherichia coli
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1803
Illegal SpeI site found at 60
Illegal PstI site found at 1938 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 19
Illegal NheI site found at 42
Illegal SpeI site found at 60
Illegal PstI site found at 1938 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1803
Illegal SpeI site found at 60
Illegal PstI site found at 1938 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1803
Illegal SpeI site found at 60
Illegal PstI site found at 1938 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Using bioinformatics tools, the best DNA gene sequence corresponding to the mRNA of the original part (GenBank: X68251.1) was found. With this, the absence of signal peptide was checked and promoter, terminator and RBS have been added. The promoter used was BBa_J23100, the RBS used was BBa_J61101 and the terminator used was BBa_B0015. The sequence was optimized for synthesis using the bacteria Escherichia coli and restriction enzymes were removed for 3A Assembly.
Source
https://www.ncbi.nlm.nih.gov/nuccore/X68251.1/
References
M.J.I. Paine, H.P. Desmond, R.D.G. Theakston, J.M. Crampton Purification, cloning and molecular characterization of a high molecular weight hemorrhagic metalloprotease, jararhagin, from Bothrops jararaca venom. Insights into the disintegrin-like gene family J. Biol. Chem., 267 (1992), pp. 22869-22876.
BJARNASON, J. B.; FOX, J. W. Snake venom metalloendopeptidases: Reprolysins. Proteolytic Enzymes: Aspartic and Metallo Peptidases, p. 345–368, 1995.
W. Bode, F.X. Gomis-Ruth, W. Stockler Astacins, serralysins, snake venom and matrix metalloproteinases exhibit identical zinc-binding environments (HEXXHXXGXXH and Met-turn) and topologies and should be grouped into a common family, the ‘metzincins’ FEBS Lett., 331 (1993), pp. 134-140.