Difference between revisions of "Part:BBa K314101"

Latest revision as of 05:03, 23 October 2010

Low constitutive expression cassette

Low constitutive protein expression insert includes f1 origin K314110, a low expression promoter J23114, 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

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
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 470
Illegal NheI site found at 493
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]

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 <partinfo>BBa_K314101 short</partinfo>
-Low constitutive protein expression insert includes f1 origin [https://parts.igem.org/Part:BBa_K314110 K314110], a high expression promoter [https://parts.igem.org/Part:BBa_J23100 J23114], 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.
+Low constitutive protein expression insert includes f1 origin [https://parts.igem.org/Part:BBa_K314110 K314110], a low expression promoter [https://parts.igem.org/Part:BBa_J23100 J23114], 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.
 ===Usage and Biology===