Part:BBa_K4686068
pMCherry+ET_Link_EGF_mEGFP
This plasmid contains the pMLB_mCherry plasmid backbone along with the heat-stable Enterotoxin ST-IA/ST-P, which is the non-virulent heat-stable enterotoxin produced by Enterotoxigenic E. coli. 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_K4686034 and BBa_K4686030 on the T7_PB_SFFV_CD4_EGFf_mEGFP_TEVs_CD28_ plasmid. The BBa_K4686034 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 Enterotoxin was sourced from UniProt.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2432
Illegal XbaI site found at 1382
Illegal PstI site found at 7935 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2432
Illegal PstI site found at 7935 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2432 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2432
Illegal XbaI site found at 1382
Illegal PstI site found at 7935 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1306
Illegal EcoRI site found at 2432
Illegal XbaI site found at 1382
Illegal PstI site found at 7935 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1613
Illegal BsaI.rc site found at 7761
Illegal SapI site found at 6678
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