We used the method of PCR and oligonucleotide ligation to succeed the construction of our UNITs, including SNAP UNIT, VIKA UNIT and mCHERRY UNIT.
Figure 1: map of SNAP UNIT
Figure 2: map of VIKA UNIT
Figure 3: map of mCHERRY UNIT
As our UNITs all contain XbaI and SpeI enzyme sites, we used 3A assembly and successfully obtained our COUNTER CIRCUIT which consists of SNAP UNIT and VIKA UNIT, and our REPORTER CIRCUIT which consists of SNAP UNIT and mCHERRY UNIT.
Figure 4: map of COUNTER CIRCUIT
Figure 5: map of REPORTER CIRCUIT
Our STARTER CIRCUIT was constructed by LR reaction that recombined recombinase Cre into the tet-on vector. Due to the complication of the tet-on system which is hard to adjust into the biobrick, and because we submitted Cre last year, this circuit has not contributed to our parts.
Figure 6: map of STARTER CIRCUIT