Team:Macquarie Australia/Protocols/Designing Gibson Assembly Fragments

From 2012.igem.org



Contents

Haemoxygenase

Original Gene Sequence: http://www.ncbi.nlm.nih.gov/nuccore/BA000022


Team mates performing codon optimisation

Deinococcus radiodurans Bacteriophytochrome

Original Gene Sequence: http://www.ncbi.nlm.nih.gov/nuccore/15807672/?from=53908&to=56175&strand=true&report=genbank

Agrobacterium tumefaciens Bacteriophytochrome

Original Gene Sequence: http://www.ncbi.nlm.nih.gov/nuccore/159184118/?from=1954292&to=1956502&strand=true&report=genbank

BioBrick assembly using Gibson method: Fragment design

  • Codon optimisation:

The Amino acid sequences for Haemoxygenase (synechocystis) and Bacteriophytochrome (Deinococcus radiodurans and Agrobacterium tumefaciens) were initially run through the Reverse Translate program (http://www.bioinformatics.org/sms2/rev_trans.html) for codon optimisation in E. coli BL21 .