IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/04/12
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Objective: Part LovTap design
We decided to synthesize a new LovTap part, that in comparison with the Part:BBa_K191003  that is already at the registry, has the following differences:
1. The 2 PstI restriction sites were removed from the coding region of LovTap.
The reason of the conformational change is the following:
Cys450 side chain is involved in light state in bond formation with the cofactor. Cys450 can assume two conformational states, called here ON and OFF, and corresponding respectively, to the Sg being near or far from FMN.
The isoleucine 427 is quite big. But not enough to push the cystein's side chain significantly toward the cofactor. So we choose to replace this ILE427 by an PHE427, an amino acid which is much bigger and have more or less the same propreties than the ILE.
atgttggctactacacttgaacgtattgagaagaactttgtcattactgacccaaggttgccagataatcccttta tattcgcgtccgatagtttcttgcagttgacagaatatagccgtgaagaaattttgggaagaaattgtcgttttctacaaggtcctgaaactgatcgcgc gacagtgagaaaaattagagatgccatagataaccaaacagaggtcactgttcagctgattaattatacaaagagtggtaaaaagttctggaacctcttt cacttgcagcctatgcgagatcagaagggagatgtccagtactttattggggttcagttggatggaactgagcatgtccgagatgctgccgagagagagg gagtcatgctgattaagaaaaccgccgaaaatattgatgaggcggcatttgtcgacctgcttaagaatg cctaccaaaacgatctccatttaccgttgttaaacctgatgctgacgccagatgagcgcgaagcgttggggactcgcgtgcgtattgtcgaagagctgtt gcgcggcgaaatgagccagcgtgagttaaaaaatgaactcggcgcgggcatcgcgacgattacgcgtggatctaacagcctgaaagccgcgcccgtggag ctgcgccagtggctggaagaggtgttgctgaaaagcgattaataa
1. Registry entry: Part:BBa_K191003
2. Wiki Team: EPF-Laussane. Simulations and results of predicted lovTAP mutations.
3. Wiki Team: EPF-Laussane. LovTAP characterization results.
4. Registry entry: Part:BBa_B0030
5. Registry entry: Part:BBa_B0032
6. Registry entry: Part:BBa_R0010