Generator

Part:BBa_K325259

Designed by: Ben Reeve and Emily Knott   Group: iGEM10_Cambridge   (2010-10-23)

Yellow Firefly Luciferase (under pBAD)
L. Cruciata
(E. coli optimised)

Input: L-Arabinose
Output: Light (yellow)

pBad/araC
I0500
mutated version of
Luciferase/LRE
K325210

This part should show similarity to BBa_K325219     Add Data       


Description
This part is based on part BBa_K325219. Two site-directed mutagenesis events have been carried out on the luciferase, an Asn286 Ser mutation was used to create part BBa_K325209 which is the green wild-type luciferase. Then a Pro452Ser mutation was used to create a yellow bioluminescence.


Colour of emission

Figure 1 - Comparison of part BBa_K325259 against other constructs created by the E.glowli team. Note that this picture should be used as a relative comparison rather than absolute colour. All images change markedly depending on the white balance used.


Compatibility
Chassis: Device has been shown to work in Top 10 (Invitrogen)
Plasmids: Device has been shown to work on pSB1C3

Assembly standards

This part is based on Part BBa_K325219. We resynthesised this part to remove three PstI restriction sites, making it compatible with BioBrick assembly standards 10 and 23.

Unfortunately, in the process of mutagenesis to create the colour change a SpeI site was introduced at site 3044. We have ordered primers to remove this site by a substitution from T->C at base 3046. BBa K325259 (planning) will be the successor to this part compatible with Assembly Standards 10 and 23. Unfortunately we did not have time to complete this before the submission deadline, therefore the part will be submitted during November 2010. In the meantime it is possible to use this part with RFC 57.

Sequences

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 3044
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal NheI site found at 3278
    Illegal SpeI site found at 3044
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2169
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 3044
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 3044
    Illegal AgeI site found at 979
    Illegal AgeI site found at 2118
    Illegal AgeI site found at 3820
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SpeI site found at 3044
[edit]
Categories
//classic/generator/uncategorized
Parameters
outputLight
positive_regulatorsL-arabinose