Difference between revisions of "Part:BBa K1943012"
 
Line 4: Line 4:
  
 
Mutated sfGFP coding sequence without KpnI recognition site
 
Mutated sfGFP coding sequence without KpnI recognition site
 +
 +
We submitted this part of super folder GFP (sfGFP) with a strong promoter <partinfo>J23116</partinfo>.
 +
 +
Under this expression system, a very bright blue-green fluorescence will be observed after 6 hours of incubation.
 +
 +
Its excited wavelength is close to the UV light.
 +
 +
we commonly manipulate it by using the UV in super clean bench.
 +
 +
The fluorescence is strong enough to be observed by naked eyes.
 +
 +
For this reason, sfGFP is always used as reporter gene under the plasmid construction.
 +
 +
This plasmid can be used as a vector.
 +
 +
After ligation, it is easy to pickup the single colony without blue-green fluorescence.
 +
 +
On the other part we submitted mutated sfGFP (msfGFP).
 +
 +
It has one bp difference with the original sfGFP on its 165th site.
 +
 +
We changed the base pair from AT to GC by using whole plasmid mutagenesis to remove KpnI restriction site.
 +
 +
We kindly hope the little effort can bring convenience to the following iGEMers.
 +
 +
In addition, we also constructed the sfGFP expression system with a terminator following the coding sequence.
 +
 +
While the two groups (with and without terminator ) showed no significant difference in our experience, we hope these parts can be provided as comparable experimental groups to quantify the efficiency of bacterial terminators.
 +
 +
[[File:Exp of sfgfp.jpeg]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 17:27, 21 October 2016


msfGFP coding sequence

Mutated sfGFP coding sequence without KpnI recognition site

We submitted this part of super folder GFP (sfGFP) with a strong promoter BBa_J23116.

Under this expression system, a very bright blue-green fluorescence will be observed after 6 hours of incubation.

Its excited wavelength is close to the UV light.

we commonly manipulate it by using the UV in super clean bench.

The fluorescence is strong enough to be observed by naked eyes.

For this reason, sfGFP is always used as reporter gene under the plasmid construction.

This plasmid can be used as a vector.

After ligation, it is easy to pickup the single colony without blue-green fluorescence.

On the other part we submitted mutated sfGFP (msfGFP).

It has one bp difference with the original sfGFP on its 165th site.

We changed the base pair from AT to GC by using whole plasmid mutagenesis to remove KpnI restriction site.

We kindly hope the little effort can bring convenience to the following iGEMers.

In addition, we also constructed the sfGFP expression system with a terminator following the coding sequence.

While the two groups (with and without terminator ) showed no significant difference in our experience, we hope these parts can be provided as comparable experimental groups to quantify the efficiency of bacterial terminators.

Exp of sfgfp.jpeg

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 13