Part:BBa_K1884009
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
Usage
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 3123
Illegal NotI site found at 3221 - 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1444
Illegal AgeI site found at 2173 - 1000INCOMPATIBLE 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
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