Device
Part:BBa_K2457004:Design
Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano Group: iGEM17_Amazonas_Brazil (2017-07-09)
Donor DNA for HDR into LacZ
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 321
Illegal NheI site found at 1164 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
It has 1810bp and was designed to contain two homologous arms with 300bp from the LacZ gene located on the Operon Lac. According to Zhao et al (2016), the use of recombination arms with 300bp increases the efficiency in 100%. On the other hand, if you have a 150bp arm, the efficiency decreases to 63%.</p>
Between the homologous arms, lies the GFP gene, flanked by NheI sites, allowing an RFC10 versatile one-step compatibility, the insertion of any other gene in GFP site.