Device

Part:BBa_K2018050

Designed by: Joel Vej-Nielsen   Group: iGEM16_SDU-Denmark   (2016-10-13)


BioBrick that produces large amounts of plastic and secretes it

This BioBrick contains phaCAB with a hybrid promoter, pnatothenate kinase and a hemolysin based type I secretion system.

Sequence and Features The BioBrick encodes the inner membrane transport complex proteins HlyB and HlyD. These transport proteins, derived from Escherichia coli strain SEC470. The BioBrick also encodes part of the hemolysin toxin, HlyA, fused with phasin, a protein that binds to PHB granules. Furthermore the BioBrick encodes phaCAB, so it is able to produce and secrete PHB.

The pantothenate kinase II is derived from staphylococcus aureus, as this protein isn't feedback inhibited by coenzyme A and coenzyme A esthers.

A proteome analysis of E. coli with and without panK revealed candidate genes that are upregulated and downregulated due to the expression of panK. Accesion numbers of downregulated genes: P0A9Q3, P05825, P09169, P0AEE6, P0ABI6, P0AAU1, E0J5J4, P0A6J9, P0AFJ3, P64469 & P0ABU1. Accesion numbers of upregulated genes: P76537, A7ZUD3, A7ZR34, P0AD62, P0C0S2, P21507, A7ZSU3, A7ZU52, P63390, P39173, P0AC71, P06612, P00805, P0ABZ8, P60753, P09831, P33602, P0AE20, A7ZJ19, A7ZPS4 & A7ZPR5.


T--SDU-Denmark--PHB1.png As compared with Top10 without the construct, the generation time is higher for cells containing K2018050.

T--SDU-Denmark--PHBOD.png The highest OD can be achieved with growth in terrific broth (TB) media.

T--SDU-Denmark--PHBOD.png Visible aggregates form in the media of bacteria containing the BioBrick


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 9108
    Illegal NheI site found at 9131
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 981
    Illegal BglII site found at 1806
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 287
    Illegal NgoMIV site found at 358
    Illegal NgoMIV site found at 958
    Illegal NgoMIV site found at 1270
    Illegal NgoMIV site found at 1549
    Illegal NgoMIV site found at 2201
    Illegal NgoMIV site found at 2223
    Illegal AgeI site found at 6620
    Illegal AgeI site found at 9518
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4067
    Illegal SapI.rc site found at 8477


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