Generator

Part:BBa_K624061

Designed by: Yi-Hong Liou   Group: iGEM11_NYMU-Taiwan   (2011-10-05)


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Safety Flag

The iGEM Safety Committee has placed a Red Flag on this part. This part presents safety risks beyond what is normal for the Registry. Researchers who plan to acquire and use this part should take special care to ensure they use it safely and responsibly. Contact safety (AT) igem (DOT) org with any questions.

Reason: Listeriolysin and Invasin parts

If you are an iGEM team, you must submit a Check-In before acquiring and using this part! See the 2017 Safety Page for more information.


pYMB essentials + RBS(trunc.) + LLO

pYMB essentials + RBS(trunc.) + LLO

pYMB (BBa_K624004) is described to contain the ori (origin of replication), rep gene (required for replication) of pMGT. Once the synthetic work had been done, pYMB is constructed by equipping the ori, the appropriate promoter for AMB-1 (Pmms16 and Pmsp3 as our candidate) and rep gene on the commercial plasmid pUG19 which was revised as the expression of Biobrick backbone pSB1A1 with promoter Pmsp1. The constructed vector is capable of replicating within both E. coli and AMB-1, fully sufficing a competent shuttle vector for genetic engineering the magnetotactic bateria.

The ribosome binding site is from Psmp3 with 6 bps truncated.(BBa_K624013).

Listeriolysin O (LLO)is a hemolysin produced by the bacterium Listeria monocytogenes, a pathogen that is responsible for listeriosis. LLO is activated within phagosomes of cells that have phagocytosed L. monocytogenes cells, and lyses the membrane of phagosome. The bacteria is then able to escape into the cytosol without damaging the plasma membrane, and grow intracellularly. This would result in the protection from extracellular immune system.


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2558
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 898
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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