Part:BBa_K101014

AraC promoter, RBS, GFP and terminator

A previously not characterized promoter was characterized using the standard promoter characterization technique. The promoter R0080 was cloned with RBS-GFP-terminator (E0240). The GFP expression was measured and found to be weak compared to promoter J61002.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 103
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 820

The device has promoter (araC regulated) which has truncated araC operator sites (Part: R0080). Prompter is cloned upstream to part E0040 (GFP). Promoter was characterized by the addition of 0, 5 and 10 mg/ml of arabinose to the growing cells. Promoter activity increased with time and found to be maximum after 3.5 hours of induction (graph is attached).

Input: arabinose (5 & 10 mg/ml)

Output: Green Fluorescence (501-520 nm)

Experiment	Characteristic	Value
Promoter Activity	Maximum Fluorescence (with 10 mg/ml) arabinose	223385.24 (Normalized fluorescence at OD₆₀₀ ~1.0) 3.5 hours post induction.
	Minimum Fluorescence (with 10 mg/ml) arabinose
	Minimum Fluorescence (with 10 mg/ml) arabinose	67986.1 (Normalized fluorescence at OD₆₀₀ ~0.4) 5 minutes post induction
Promoter Activity	Maximum Fluorescence (with 5 mg/ml) arabinose	206750.5(Normalized fluorescence at OD₆₀₀~1.0)3.5 hours after the growth.
Promoter Activity	Minimum Fluorescence (with 5 mg/ml) arabinose	67126.57 (Normalized fluorescence at OD₆₀₀~0.4)5 minutes after the growth.
Growth of culture in MOPs (minimal) media	Doubling time of the recombinant cells	~ 55 Minutes
Response time (after addition of arabinose	~30Minutes

Compatibility
Chassis: Device has been shown to work in BBa_E0040, Bba_B0010, and Bba_B0012

Plasmids: Device has been shown to work in pSB3K3