Part:BBa_K4686070
pMCherry+O33434_Link_EGF_mEGFP
This plasmid contains the pMLB_mCherry plasmid backbone along with the L-cysteine S-thiosulfotransferase subunit SoxA (O33434) signal peptide. This signal peptide is linked to an EGF ligand bound to mEGFP via a GGSG linker. As a signal peptide, this protein contains a signal sequence that will interact with signal recognition particles to then direct ribosomes to the endoplasmic reticulum for co-translational insertion to take place.
Used primers BBa_K4686027 and BBa_K4686028 on the pMLB_mCherry plasmid backbone, and primers BBa_K4686036 and BBa_K4686030 on the T7_PB_SFFV_CD4_EGFf_mEGFP_TEVs_CD28_ plasmid. The BBa_K4686036 primer contained the signal peptide sequence sourced from UniProt, so the resulting insert contains the EGF ligand sequence from T7 as well as the signal peptide sequence. The pMLB_mCherry_GGBsMTS plasmid backbone was sourced from Piet de Boer. The T7 plasmid was sourced from our mentor Beatrice Ramm. The sequence for the L-cysteine S-thiosulfotransferase subunit SoxA (O33434) was sourced from UniProt.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2456
Illegal XbaI site found at 1382
Illegal PstI site found at 7959 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2456
Illegal PstI site found at 7959 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2456 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2456
Illegal XbaI site found at 1382
Illegal PstI site found at 7959 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2456
Illegal XbaI site found at 1382
Illegal PstI site found at 7959 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1637
Illegal BsaI.rc site found at 7785
Illegal SapI site found at 6702
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