Composite

Part:BBa_K1985015

Designed by: Hanya Benotmane   Group: iGEM16_Kent   (2016-10-14)
Revision as of 15:27, 17 October 2016 by JJECornish (Talk | contribs) (→‎Validation)


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE 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.

File:Filename.jpeg
Figure 1. Agarose gel of the restriction digest of BBa_K1985012 in pSB1A3, with EcoRI and PstI.



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