Part:BBa_K2717020
final1(GFP-TGATG-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:
By ligating the stop codon of GFP with the initiation codon of pchBA to form the structure of TGATG, the coupled expression of GFP and pchBA is realized, 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 1023
Illegal NgoMIV site found at 1427
Illegal NgoMIV site found at 1552
Illegal NgoMIV site found at 1563
Illegal NgoMIV site found at 1844
Illegal NgoMIV site found at 2312
Illegal AgeI site found at 4537
Illegal AgeI site found at 5429 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5411
Illegal BsaI.rc site found at 644
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