Part:BBa_K4375018
MiniNano for bacterial surface display
The MiniNano is a compact construct meant for extracellular protein (like tumour-specific receptor, antigen, etc.) detection via fluorescence. This protein has 3 domains. (1) INPNC surface display module (INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringae, and it is used for displaying proteins on bacteria's outer membrane (BBa_K4375007)). It is linked to (by a flexible GS-linker, BBa_K4375010) ) (2) a small, near-infrared emitting fluorescent protein, miRFP670 nano (BBa_K4375008) ), which is connected to an anti-EGFR specific Nanobody, 7D12, via llama-derived igA hinge region linker (BBa_K4375011). Moreover, it contains a Myc epitope tag, so the specific interaction can be detected in another way. This complex detection device is expressed under the constitutive promoter J23101* (BBa_K4375003) The Nanobody in the construct can be easily swapped with other protein-domains, as its sequence is framed by BamHI and SphI restriction sites.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1055
Illegal PstI site found at 1313 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 29
Illegal PstI site found at 1055
Illegal PstI site found at 1313 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1055
Illegal PstI site found at 1313 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1055
Illegal PstI site found at 1313
Illegal NgoMIV site found at 118
Illegal NgoMIV site found at 451
Illegal AgeI site found at 869
Illegal AgeI site found at 1523
Illegal AgeI site found at 1655 - 1000COMPATIBLE WITH RFC[1000]
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