Coding

Part:BBa_K1499502:Design

Designed by: Jovita Byemerwa   Group: iGEM14_StanfordBrownSpelman   (2014-09-30)


Cellulose Acetate Esterase (Cae)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 1533
    Illegal EcoRI site found at 25
    Illegal XbaI site found at 40
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 25
    Illegal NheI site found at 1384
    Illegal SpeI site found at 1534
    Illegal PstI site found at 1548
    Illegal NotI site found at 31
    Illegal NotI site found at 1541
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 25
    Illegal BamHI site found at 625
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 25
    Illegal suffix found in sequence at 1534
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 25
    Illegal XbaI site found at 40
    Illegal SpeI site found at 1534
    Illegal PstI site found at 1548
    Illegal NgoMIV site found at 588
    Illegal AgeI site found at 326
    Illegal AgeI site found at 499
    Illegal AgeI site found at 730
    Illegal AgeI site found at 912
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Neisseria Sicca secretes the enzyme extracellularly and hence it has a signal sequence to aid for secretion. In order to use the gene in E.coli, we removed the signal sequence of Neisseria Sicca and added an E.coli specific secretion tag. In addition to that, we added a his-tag at the c'terminus end of the protein to aid purification of the protein.


Source

Genomic sequence of Neisseria Sicca

References