Coding
Part:BBa_K1111008:Design
Designed by: Luisa Spisak, Marcelle Isaline Arrigo Group: iGEM13_EPF_Lausanne (2013-09-19)
Gelatinase E (GelE)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1885
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1885
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1418
Illegal BglII site found at 1469
Illegal BamHI site found at 1812 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1885
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1885
Illegal NgoMIV site found at 1114
Illegal AgeI site found at 1487 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 301
Illegal BsaI.rc site found at 1018
Illegal SapI.rc site found at 1953
Design Notes
The coding sequence (CDS) was obtained by direct PCR of the E. faecalis genomic DNA, that we ordered to prevent any safety issues since this bacterium is of type 2. A His tag was also added at the beginning of the protein CDS and a linker at the 3' end.
Link for the NCBI GelE CDS:http://www.ncbi.nlm.nih.gov/nuccore/D85393.1
GelE forward primer:5'-ATG CACCACCACCACCACCAC AAG GGA AAT AAA ATT TTA TAC CAT TTT A-3'
GelE reverse primer:5'-ACC ACC AGA TTG AAA ATA CAA ATT TTC ACC TTC ATT GAC CAG AAC AGA TTC-3'
PCR Results:
Source
E.faecalis