Part:BBa_K737040
galK::GFP generator ligated to TetR generator and constitutive sRNA device
Small RNA Spot42 controls the GFP mRNA translation by blocking the RBS in galK.The galK and E0040 is fused together using ClaI site to avoid stop condon in standard scar.J23106 is a moderate promotor,and galK’s RBS is a moderate one.
[J23106,P0440]is a TetR generator, which represses tet promotor and can be induced by aTc. aTc concentration gradient results in different sRNA Spot42 levels in cells when presented R0040, which can be monitored by our galK::GFP generator,where galK is the RBS of GFP and the target for Spot42. This device was mistaken for IPTG inducible sRNA device originally due to some ligation mistake,but we actually found this one is perfect to express full level sRNA to verify sRNA function without induction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 1078
Illegal NheI site found at 1101 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1890
In the picture,L3 represents BBa_ K737040,JgE represents BBa_K737034,DsrA is control.
[J23106,P0440]is a TetR generator, which represses tet promotor and can be induced by aTc. aTc concentration gradient results in different sRNA Spot42 levels in cells when presented R0040, which can be monitored by our galK::GFP generator,where galK is the RBS of GFP and the target for Spot42. This device was mistaken for IPTG inducible sRNA device originally due to some ligation mistake,but we actually found this one is perfect to express full level sRNA to verify sRNA function without induction.
| n/a | galK::GFP generator ligated to TetR generator and constitutive sRNA device |