Part:BBa_K2623030
A composite part of OmpA-SpyTag and SpyCatcher-L7Ae-mRFP1
Summary
This part is a composite part constructed by linking BBa_K2623024 and BBa_K2623025 together. BBa_K2623024 can express OmpA inserted with SpyTag (ST) at its C-termini. BBa_K2623025 can express archaeal ribosomal protein protein L7Ae inserted with SpyCatcher (SC) at its N-termini and mRFP1 at its C-termini under the induction of IPTG at 0.5 mM concentration.
Usage and Biology
Though we have revealed the OmpA-ST inside OMVs (BBa_K2623021), we are inclined to figure out the ST/SC bioconjugation inside OMVs. Once IPTG is added, L7Ae could be expressed with SpyCatcher and RFP modification and then be encapsulated into OMVs through ST/SC conjugation. We also constructed another form of BBa_K2623030 (NG9-1)-BBa_K2623028 (NG8-1).The only difference is that the latter part links BBa_K2623022 (NG2) instead of NG4 with NG5 (Figure 1). Through the fluorescence intensity of mRFP1, we can compare and evaluate the efficiency of SpyTag/SpyCather bioconjugation inside OMVs . Then the better one would be uesd to construct our final gene circuit-BBa_K2623032.
 Figure 1. a) Genetic circuit of SC-L7Ae-RFP. b) The spectrogram of mRFP1. Reference: https://www.semrock.com/ |
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 1965
Illegal NheI site found at 1988 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 65
Illegal AgeI site found at 3122
Illegal AgeI site found at 3234 - 1000COMPATIBLE WITH RFC[1000]
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