Plasmid_Backbone
Part:BBa_K2448036:Design
Designed by: jeremy armetta Group: iGEM17_Evry_Paris-Saclay (2017-10-27)
pSB1C3 BsmBI free
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2049
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 2055 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2049
Illegal XhoI site found at 1033
Illegal XhoI site found at 1925 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 2049
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 2049
Plasmid lacks a suffix.
Illegal XbaI site found at 2064
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Design Notes
To disrupt the BsmBI recognition site CGTCTC by site-directed mutagenesis we choose to change its sequence to CGTGTC. This site is present in the chloramphenicol resistance gene so care was taken that the mutation should not affect the amino acid composition of the protein. The chosen mutation is a synonymous one GTC(Val) to GTG(Val).
>BsmBI | pSB1C3: 1390 - TTC GTC TCA - 1382 F V S BBa_K2448036: 1390 - TTC GTG TCA - 1382 F V S
Source
site directed mutagenesis on the pSB1C3 bearing the BBa_J04450 reporter.
References
[1] Engler C, Kandzia R, Marillonnet S. A one pot, one step, precision cloning method with high throughput capability. PLoS One (2008) 3, e3647.