Part:BBa_K3285000
Blue Light Repressible Promoter
This composite promoter consists of 6 basic parts that include 2 promoters, 3 coding sequences and one double terminator.
USAGE AND BIOLOGY
There is a constitutive promoter that constitutively expresses the yf1 and fixJ genes. YF1 is a fusion protein made of YtvA (Light sensitive protein) and FixL (phosphorylates FixJ). The phosphorylated FixJ induces pFixK2 promoter and hence mrfp is expressed. As a result the RFP is constitutively expressed when there is no blue light. Now, in the presence of blue light, the conformation of Ytv1 changes hindering the phosphorylation of FixJ by FixL leading to a reduced expression of mrfp gene. Increasing the intensity of blue light further hinders this interaction leading to an even lesser expression of mrfp gene. As a result, with increasing intensity of blue light, RFP production reduces. We want to use this promoter to regulate the level of mutD5 (codes for the faulty epsilon subunit of DNA polymerase III of bacteria) in the cell thereby resulting in different levels of mutation rates in the bacteria. Our project aims at developing a tool for attaining tunability of mutation rates in E.coli using an external physical inducer (such as blue light) which will help in performing directed evolution faster and more easily.
SEQUENCE AND FEATURES
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 605
Illegal NgoMIV site found at 677
Illegal NgoMIV site found at 767
Illegal NgoMIV site found at 785
Illegal NgoMIV site found at 1297
Illegal NgoMIV site found at 1590
Illegal NgoMIV site found at 1684
Illegal AgeI site found at 319
Illegal AgeI site found at 1465
Illegal AgeI site found at 2815
Illegal AgeI site found at 2927 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1354
Illegal BsaI.rc site found at 218
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