Coding

Part:BBa_K322119:Design

Designed by: Meng Lu   Group: iGEM10_Edinburgh   (2010-10-27)


CcaS upstream + PhoR downstream


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1235
    Illegal SpeI site found at 1330
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1235
    Illegal NheI site found at 606
    Illegal SpeI site found at 1330
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1235
    Illegal XhoI site found at 1543
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1235
    Illegal SpeI site found at 1330
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1235
    Illegal SpeI site found at 1330
    Illegal AgeI site found at 1270
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Both proteins have a sensing domain and PAS domains in the external part of the protein, and HisKA and HATPase_c domains on the cytoplasmic side. The fusion site was chosen in the middle of the region between these outer and inner domains, by amplifying the required regions by PCR. An HolI site was added downstream of the upper CcaS region, and upstream of the lower PhoR region. HolI was chosen because the fusion scar would not introduce new amino acid, in a region that was very similar in CcaS and PhoR.



Source

CcaS: Synechocystis sp. (strain PCC 6803)- Synthesised by Unam Mexico 2010. PhoR: Escherichia coli (strain K12)genomic DNA.


References