Composite

Part:BBa_K1520022

Designed by: yue wu   Group: iGEM14_Nanjing-China   (2014-09-22)
Revision as of 01:28, 18 October 2014 by Sagyman (Talk | contribs)

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Pcons2-rbs-lacI-Ter-Plac-rbs-luxI-Ter-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-Ter

Plasmid in positive feedback verification: Pcons2-rbs-lacI-Ter-Plac-rbs-luxI-Ter-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-Ter It is a positive feedback plasmid using luxI as key to feedback. Introducing IPTG will produce rfp and luxI, and luxI will work back to enhance production of rfp.

Positive feedback.jpg

As is shown in the picture, plasmid with “LuxI-T”(purple) shows higher fluorescence intensity than the control(blue), so this kind of plasmid can be more efficient than the control, however, it is not as efficient as the plasmid with “LuxR-LuxI-T”(Yellow).

K1520021.jpg

From left to right, Marker(1Kb),

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-Ter,

Pcons2-rbs-lacI-T-Plac-rbs-luxI-T-Pcons2-rbs-luxR-Ter-Prlux-rbs-rfp-rbs-luxI-rbs-luxR-Ter.

The substances of electrophoresis are made from PCR of plasmid with VF2 and VR as their primers.

Sequence and Features


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 2380
    Illegal NheI site found at 2403
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1208
    Illegal BglII site found at 2222
    Illegal BglII site found at 4818
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3358


[edit]
Categories
//function/biosynthesis/ahl
//function/cellsignalling
Parameters
biology
controlluxR, HSL
directionForward
ec_numnone
functionMixed
keggnone
ligandsHSL
n/aconstitutive promoter family member
negative_regulators
o_h
o_l
positive_regulators1
proteinluxI-LVA
swissproP12747
tagLVA
uniprotP48445