Difference between revisions of "Part:BBa K642007"
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This is the distal region of the mid length Adh1 promoter (BBa_K319005) submitted last year by the uOttawa team fused to the proximal region of the Gal10/1 promoter. It is a constitutively expressing chimeric promoter. It is also tagged with the reporter yeGFP (BBa_K319039) which was submitted by the uOttawa team last year.
 
This is the distal region of the mid length Adh1 promoter (BBa_K319005) submitted last year by the uOttawa team fused to the proximal region of the Gal10/1 promoter. It is a constitutively expressing chimeric promoter. It is also tagged with the reporter yeGFP (BBa_K319039) which was submitted by the uOttawa team last year.
  
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===Characterization of BBa_K642007===
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===Usage and Biology===
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We were able to characterize the expression of the Adh1 distal-Gal10/1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 locus of a BY4742 strain of S. cerevisiae. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for GFP fluorescence on a Cyan ADP flow cytometer.
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[[Image:C07.jpg]]
  
 
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Revision as of 17:39, 24 October 2011

Constitutive Adh1 distal-Gal1 proximal promoter tagged with yeGFP

This is the distal region of the mid length Adh1 promoter (BBa_K319005) submitted last year by the uOttawa team fused to the proximal region of the Gal10/1 promoter. It is a constitutively expressing chimeric promoter. It is also tagged with the reporter yeGFP (BBa_K319039) which was submitted by the uOttawa team last year.

Characterization of BBa_K642007

We were able to characterize the expression of the Adh1 distal-Gal10/1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 locus of a BY4742 strain of S. cerevisiae. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for GFP fluorescence on a Cyan ADP flow cytometer.

File:C07.jpg

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 BsaI site found at 89
    Illegal BsaI.rc site found at 1379