Part:BBa_K1985015
pSB1A3-[AraC-pBAD]-[CsgA-SS-Sup35NM]
This part is an improved version of biobrick (BBa_K1739002), which was designed by the Kent 2015 iGEM team. The promoter BBa_J23104 has been removed and instead an inducible arabinose promoter, part BBa_K1321333, has been used. The second part of this composite biobrick contains 2 segments: the CsgA signal sequence and Sup35NM.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Usage and Biology
For more information on biology and usage of Sup35NM with the CsgA signal sequence, see part BBa_K1739002. For information on the promoter see BBa_K1321333.
Validation
The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: xxxx for the plasmid backbone and xxx for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced.
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