Device

Part:BBa_K2457004:Design

Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano   Group: iGEM17_Amazonas_Brazil   (2017-07-09)
Revision as of 01:36, 2 November 2017 by Giomaklouf (Talk | contribs) (→‎Design Notes)


Donor DNA for HDR into LacZ


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 321
    Illegal NheI site found at 1164
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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.

Source

Mudar

References