Composite

Part:BBa_K1884009

Designed by: Changxing Hu   Group: iGEM16_SZU-China   (2016-10-11)
Revision as of 09:52, 15 October 2016 by Nllenwalker (Talk | contribs) (→‎Biology)


PSAD Promoter+GalAD-CRY2+UAS+FNR(fumarate and nitrate reduction)+PSAD Terminator

The device we constructed this year is one of the functional plasmids in the light-mediated expression system,followed the example of yeast two-hybrid system. With BD-CIB1 which is constructed in the other device(BBa_K1884007), light-mediated expression system, followed the example of yeast two-hybrid system, will be activated and starting to transcript Ferredoxin-NADP(+)Reductase(BBa_K1884006) which is on the 3 prime end of Upstream activating sequence(BBa_K1884004) in green algae

Biology

PASD promoter(BBa_K1547005) is a plant promoter from the green algae Chlamydomonas reinhardtii. It is a high expression promoter that encodes for a ferrodoxin-binding protein of photosystem I.This year we decided to built the light-mediate-controlled system in green algae Chlamydomonas reinhardtii.Based on the origin of PSAD promoter, we thought it would be fully competible in our project.

AD-CRY2, a fusion protein, is for use in a yeast-two-hybrid system, and a VP16 DNA activating domian fused to its N terminus.In order to control DNA transcription by blue light, the system is based on a two-hybrid interaction in which a light-mediated protein brings together two halves of a split transcription factor. CRY2 will disconnected with CIB1 in the dark and halt the DNA transcription

UAS

FNR

PSAD terminator is a plant specific Terminator, and serves to increase expression of a gene placed upstream. It works in collaboration with PSAD promoter(BBa_K1547005). Both of these parts are activated by sunlight.

Usage

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3123
    Illegal NotI site found at 3221
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1560
    Illegal BglII site found at 2019
    Illegal BamHI site found at 2498
    Illegal XhoI site found at 3052
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1444
    Illegal AgeI site found at 2173
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1796
    Illegal BsaI.rc site found at 1205
    Illegal SapI.rc site found at 1313
    Illegal SapI.rc site found at 3384


[edit]
Categories
Parameters
None