Basic Parts :
In this section we present our basic parts. First we have the ones used to make our Composite parts:
- ➟ LacI-LVA wich is the LacI protein with an LVA tag causing an unstabillity of LacI that can be used in order
to have a more sensitive regulator, the codon usage was optimized for B. subtilis. - ➟ PiBlue wich is a blue pigment used as reporter in our project. The codon usage was optimized for B. subtilis.
- ➟ PFur is a Promoter Operator composed by PliaG (constitutive promoter), the FurBox wich is the DNA binding site
for the Iron metabolic pathway regulator Fur and a RBS for B. subtilis. - ➟ PLacO is a Promoter Operator composed by PlepA (constitutive promoter), two LacI binding sites and a RBS for B. subtilis.
- ➟ Terminator wich is a dual terminator for E. coli usable in B. subtilis.
All those parts were designed with specific restriction site at both ends in order to facilitate the manipulation of the biobricks.
In order to improve our detection system we also designed Operator biobricks that can be easily swaped.
- ➟ ZinOp wich is the CzrA binding site. Binding is repressed in the presence of Zinc.
- ➟ FruOp wich is the FruR binding site. Binding is repressed in the presence of Fructose.
- ➟ SucOp wich is the DegU binding site. Binding is repressed in the presence of Sucrose.
- ➟ CadOp wich is the MntR binding site. Binding is repressed in the presence of Cadmium.
- ➟ ArsOp wich is the ArsR binding site. Binding is repressed in the presence of Arsenic.
We believe that our system may be used by future iGEM teams as mean of detection for multiple parameters.