Difference between revisions of "Part:BBa K314104"
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Lac induced protein expression insert includes f1 origin K314110, Lac I generator K314111, a T7 promoter k314112, and the Elowitz standard RBS B0034.  When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.  
 
Lac induced protein expression insert includes f1 origin K314110, Lac I generator K314111, a T7 promoter k314112, and the Elowitz standard RBS B0034.  When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.  
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Lac induced T7 promoter, <partinfo>BBa_K314112</partinfo>, protein expression insert includes f1 origin [https://parts.igem.org/Part:BBa_K314110 K314110], a Lac I generator [https://parts.igem.org/Part:BBa_K314111 K314111],  a high expression promoter [https://parts.igem.org/Part:BBa_J23100 J23100], and the  Elowitz standard RBS [https://parts.igem.org/Part:BBa_B0034 B0034].  When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.
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===Usage and Biology===
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{| cellpadding="1" cellspacing="8"
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|[[Image:Washington2010_vector_assay_data.png | 650px]]
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|<partinfo>E0040</partinfo> (GFP) was inserted between S and P using standard BioBrick assembly.  The resulting constructs were grow in DH5a (constitutive promoters <partinfo>BBa_J23100</partinfo> or <partinfo>BBa_J23114</partinfo>, as well as inducible promoter <partinfo>R0011</partinfo>) or BL21(DE3)* (T7 promoter, <partinfo>BBa_K314104</partinfo>) cells overnight at 298K in either the presence or absence of 0.5mM IPTG.  The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer (ex. 485, em. 525) and normalized to cell density (OD600).  Cells that did not contain GFP were used as a blank.  As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 04:42, 23 October 2010

T7 expression cassette

Lac induced protein expression insert includes f1 origin K314110, Lac I generator K314111, a T7 promoter k314112, and the Elowitz standard RBS B0034. When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.


Lac induced T7 promoter, BBa_K314112, protein expression insert includes f1 origin K314110, a Lac I generator K314111, a high expression promoter J23100, and the Elowitz standard RBS B0034. When ligated with a protein coding sequence at the SpeI site the scar site created will create proper spacing for protein expression.

Usage and Biology

Washington2010 vector assay data.png BBa_E0040 (GFP) was inserted between S and P using standard BioBrick assembly. The resulting constructs were grow in DH5a (constitutive promoters BBa_J23100 or BBa_J23114, as well as inducible promoter BBa_R0011) or BL21(DE3)* (T7 promoter, BBa_K314104) cells overnight at 298K in either the presence or absence of 0.5mM IPTG. The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer (ex. 485, em. 525) and normalized to cell density (OD600). Cells that did not contain GFP were used as a blank. As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 126
  • 1000
    COMPATIBLE WITH RFC[1000]