<h1id="2"class="anchor">New Documentation for BBa_K3946023</h1>
<p>We characterized the part BBa_K3946023 and provided details about it's usage, as it had been previously submitted to the parts repository by the 2021 Edinburgh iGEM Team, but had not been documented thoroughly. We characterized this part by assessing the PET degradation activity with different silica tags, compared to other PETases. We showed that this part does degrade PET, but not as effectively as other parts, for example the <ahref="http://parts.igem.org/Part:BBa_K4390073">FAST-PETase</a>.</p>
<p>Beyond that we also created the DouPETase construct with different N-terminal Silica tag: Car9-DouPETase (<ahref="http://parts.igem.org/Part:BBa_K4390085">BBa_K4390085</a>) and L2NC-linker-DouPETase (<ahref="http://parts.igem.org/Part:BBa_K4390119">BBa_K4390119</a>). And we also help 2021 OG Edinburgh team to fill in more data about silica tags (L2NC,L2NC+linker, Car9+linker).</p>
<p>For any future team that plans working on the JUMP assembly, please remember to add LacO between promoter and ribosome binding site. We didn’t add the LacO and that resulted in uninducable protein expression, which made us obtain fewer proteins than expectation. Also, to avoid the error in JUMP assembly procedures, please dilute all the Lv.0 parts to 20 fmol/µl, therefore, you can take out 1 µl from every part you needed to make sure the correct amount of Lv.0 plasmids are added into the reaction. </p>
<p>To avoid the error in JUMP assembly procedures, please dilute all the Lv.0 parts to 20 fmol/µl, therefore, you can take out 1 µl from every part you needed to make sure the correct amount of Lv.0 plasmids are added into the reaction. </p>
<p>For any future team that plan to immobilize enzymes and aptamers on the silica beads, please keep in mind that too much enzymes on the same silica bead would inhibit the enzyme activity overall, since the surface area on each silica bead is limited and can’t support infinite enzymes. So, remember to check the amount of protein loading on each silica bead to make sure enzymes on the same bead are not crowded. </p>
