Part:BBa_K2717021
final2(GFP-hairpin-pchBA-terminator-emrr-RBS2000-GDH-v5 tag-his tag)
The main function of this vector is to screen the strains capable of high-yielding the target gene GFP through a positive feedback regulation system on the vector,
function:
Coupling expression of GFP and pchBA was achieved by ligating GFP behind the hairpin structure and ligating pchBA gene in the back, that is, when RNA polymerase is translated into the stop codon of GFP, there is a certain probability to directly initiate the translation of pchBA. And pchBA is a key enzyme capable of producing salicylic acid in cells, which can catalyze the production of salicylic acid. At this time, the promoter of the gdh gene on the vector, emrR binding promomter, was inhibited by the repressor protein produced by the emrR gene carried on the E. coli genome, so the gdh gene could not be expressed smoothly. And the emrR repressor protein will detach in the presence of salicylic acid, which makes gdh smoothly express, that is, the yield of GFP increased with the increase of pchBA expression, the repressive effect of emrR binding promoter decreased with the increase of salicylic acid, the expression of gdh increased with the increase of salicylic acid, and the high-yield bacteria obtains the growth advantage, which forms a positive feedback adjustment.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1070
Illegal NgoMIV site found at 1474
Illegal NgoMIV site found at 1599
Illegal NgoMIV site found at 1610
Illegal NgoMIV site found at 1891
Illegal NgoMIV site found at 2359
Illegal AgeI site found at 4584
Illegal AgeI site found at 5476 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5458
Illegal BsaI.rc site found at 644
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