Part:BBa_M31332
M13 Refactoring
I attempted to refactor the M13K07 phage by removing the overlapping areas of genes between the HpaI site of gene 2 and the BamHI site of gene 3. I did this by duplicating each gene (promoter, rbs, and ORF) and placing it after the ORF of the previous gene. I mutated the RBS's of each overlapping area to a silent mutation to prevent readout of the overlapping sections. There were 6 places of overlap, and I have included what changes I made. As a result, I have added 769 bp to the original section of DNA. In between each gene, I left 11 extra base pairs in order to introduce restriction sites so that each part can be manipulated individually. I want to see what others have done before attempting to introduce new restriction endonuclease sites. I made 4 different basic parts and joined them to together to make the composite M13.1.
| Overlapping Areas | Original Sequence | Modified sequence |
|---|---|---|
| Gene 2 ORF and Gene 10 RBS | At bp 478: gcatttgag | gcGtttgag |
| Gene 2 ORF and Gene 5 RBS | At bp 826: gcataaggt | gcGtaaggt |
| Gene 10 ORF and Gene 5 RBS | At bp 1286: gcataaggt | gcGtaaggt |
| Gene 5 ORF and Gene 7 RBS | At bp 1609: gttccggct | gtCccggct |
| Gene 7 ORF and Gene 9 RBS | At bp 1733: cgctggggg | cgAtggggg |
| Gene 9 ORF and Gene 8 RBS | At bp 1869: aatggaaac | aaCggaaac |
The refactored genome is Part BBa_M31332 on the Registry of Standard Biological Parts
| None |