Composite

Part:BBa_K4229066

Designed by: Nikita Edel   Group: iGEM22_Freiburg   (2022-09-30)
Revision as of 06:59, 12 October 2022 by NikitaEdel (Talk | contribs)


mVenus with N-terminal spyCatcher regulated by tetA/B promotor

mVenus2 is with mTurqouise2(BBa_K4229067) one of our reporter genes, with which we tested the building and catching of the minimal(BBa_K4229047) and full wiffleball(BBa_K4229049) and SPD5 (BBa_K4229078).

The snoop/spyTag snoop/spyCatcher was tested with those two reporter genes. You can see a fluorescent signal in the bacteria that expressed either of those two. You can see foci (especially well with the full wiffleball) for both of those reporter genes:

Fluorescent microscopy of T1 catching the mVenus2 and mturquoise2 when the minimal or full BMC construct is expressed: A Controls for induction; B T1 with and without the Spy/Snp tags; scalebar 5µm





























It was later confirmed that in fact mVenus was cached by T1 by a western blot comparing the full wiffleball with and without the necessary tags.

Western Blot comparison of the BMC minimal wiffleball with and w/o tags (pHT1) + mVenus2






















Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 59
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 59
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 59
    Illegal BglII site found at 68
    Illegal XhoI site found at 1075
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 59
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 59
    Illegal AgeI site found at 508
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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