Coding

Part:BBa_K1111008:Design

Designed by: Luisa Spisak, Marcelle Isaline Arrigo   Group: iGEM13_EPF_Lausanne   (2013-09-19)

Gelatinase E (GelE)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1885
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1885
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1418
    Illegal BglII site found at 1469
    Illegal BamHI site found at 1812
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1885
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1885
    Illegal NgoMIV site found at 1114
    Illegal AgeI site found at 1487
  • 1000
    INCOMPATIBLE 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:

Figure 2: PCR: GelE insert























Source

E.faecalis

References