DNA

Part:BBa_M31332

Designed by: Andrew Ji   Group: BE.109 Instructors BE.109 2006 MIT 20.109 Spring07 MIT 20.109 Fall07   (2007-02-28)

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

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