Supplementary info

Guide for the Supplementary Information:

• S.1 Detailed JUMP explanations
  A detailed table of parts used for our DNA assemblies.
• S.2. DNA sequences of all parts
  A detailed table of parts used for our DNA assemblies.
• S.3 All Plates Labelled
  A list of all the DNA sequences we used in the project.
• S.4 SDS page labelled
  A confirmation of MT expression in 2 species and their WT and mutant constructs.
• S.5 Colony PCRs labelled
  A confirmation of inserted DNA and the assembly for all our constructs.
• S.6 Mutation Sites
  The proposed mutation sites for all Metallothionein species found using MSA done phylogenetically similar MTs.
• S.7 Site Specific Mutagenesis
  List of all mutagenic primers for GeneOrator along with explanation.
• S.8 Targeted Mutagenesis primers
  List of all mutagenic primers for Targeted mutagenesis along with explanation.
• S.9 Generalised PS1 and PS2 primers
  The sequence of the primers used for colony PCR and Error-prone PCR.
• S.10 RT-DNA to ORF.py
  A python script for the conversion Reverse transcription sequences into Open Reading Frames.
• S.11 clean_automation.sh
  A bash shell script for the automation of the docking simulations run on AutoDock 4.2.
• S.12 Summarised Docking statistics
  The pass/fail metric for each docked metallothionein’s cysteine residues and the free energy per ion data.
• S.13 Docking Log Summaries of the best model
  Detailed logs of the raw data generated per cysteine of each MT.
• S.14 Reagent List
  A list of the reagents used by us.

Members of this iDEC team also participated in the iGEM 2022 competition. A lot of elements of this project were adapted to the iGEM wiki, for more information on our iGEM project click here.