Composite

Part:BBa_K3187008

Designed by: iGEM TU_Darmstadt 2019   Group: iGEM19_TU_Darmstadt   (2019-10-12)


GGGG-Tag for Sortase-mediated Ligation x mCherry Fluorescence Protein

Profile

Name GGGG-mCherry
Base pairs 1074
Molecular weight 28.5 kDa
Origin synthetic, derived from Discosoma sp.
Parts mCherry, GGGG-sequence, T7 Promoter, lac Operator, GASPAG Linker, Strep-Tag II, T7 Terminator
Properties Red fluorescent, Ex λ: 587nm, Em λ: 610 nm

Usage and Biology

mCherry (BBa_K3187026) is a red fluorescent protein. Which is a synthetic protein derived from Discosoma sp. by directed evolution. The N-terminal GGGG-sequence (BBa_K3187018) can be fused to a protein with a C-terminal LPETGG-Sortase A link (BBa_K3187019) by Sortase A. We use mCherry as an easily imaged reporter for checking if the coupling worked.
The coding sequence was cloned in pET24 vector, containing the sequence of mCherry, a GGGG-sequence, a GASPAG Linker (BBa_K3187038), a Strep-Tag II (BBa_K3187025) for Purification, a T7 Promoter with lac Operator and an RBS (BBa_K3187029), a T7 Terminator (BBa_K3187032), a Start-Codon (BBa_J70593) and a Stop-Codon (BBa_K2868029). The coding sequence consists of 851 bp which are translated to 260 amino acids.[1]

Results

For results regarding this part, please have a look at (BBa_K3187028)


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 854
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 854
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 915
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 854
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 854
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None