Difference between revisions of "Part:BBa K2123204:Design"
Mctastolfi (Talk | contribs) |
|||
Line 6: | Line 6: | ||
− | ===Design Notes=== | + | ===Design Notes: In silico study=== |
− | + | ||
+ | To construct our final bioaccumulation device, we followed the steps below: | ||
+ | <b>1st Step: </b> | ||
− | + | <center>https://static.igem.org/mediawiki/2016/thumb/2/23/UFAM_UEA_FTQ_DESIGN_2.png/800px-UFAM_UEA_FTQ_DESIGN_2.png"</center> | |
− | + | Phytochelatin digestion with EcoRI + SpeI releasing Lpp-OmpA with NdeI and XbaI. To linearize Lpp-OmpA (2526 bp) and release the promoter (~83 bp). | |
− | + | <b>3rd Step: </b> | |
+ | |||
+ | <center>https://static.igem.org/mediawiki/2016/thumb/f/fa/UFAM_UEA_FTQ_DESIGN_3.png/800px-UFAM_UEA_FTQ_DESIGN_3.png</center. | ||
+ | |||
+ | Digestion of first step construction, resulting the PO construction (JK26 promoter + Lpp-OmpA) digested with EcoRI + SpeI, releasing a 564 bp insert. The PT (Phytochelatin + Terminator) construction was linearized with EcoRI + XbaI, releasing insert of approximately 2317 bp. | ||
+ | |||
+ | Finally we had our bioaccumulator construction!! |
Latest revision as of 10:17, 22 October 2016
Bioaccumulator device: Strong promoter + OmpA fused to Synthetic Phytochelatin + B0015
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1493
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1091
Illegal NgoMIV site found at 1665
Illegal NgoMIV site found at 2902
Illegal NgoMIV site found at 2964 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1084
Design Notes: In silico study
To construct our final bioaccumulation device, we followed the steps below:
1st Step:
Phytochelatin digestion with EcoRI + SpeI releasing Lpp-OmpA with NdeI and XbaI. To linearize Lpp-OmpA (2526 bp) and release the promoter (~83 bp).
3rd Step:
Digestion of first step construction, resulting the PO construction (JK26 promoter + Lpp-OmpA) digested with EcoRI + SpeI, releasing a 564 bp insert. The PT (Phytochelatin + Terminator) construction was linearized with EcoRI + XbaI, releasing insert of approximately 2317 bp.
Finally we had our bioaccumulator construction!!