Part:BBa_K101005
Construct with LacI/LAMBDAcI promoter, RBS, GFP, Terminator
Has a promoter which is dually repressed by LacI and LAMBDAcI. The RBS is the ribosomal binding site. The GFP is a green fluorescent protein, which is a reporter gene. The Terminator is used to terminate RNA polymerase's transcription.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 787
The device has a dual promoter which is repressed by lacI and lambda CI. The input is IPTG and output is fluorescence as gene for GFP is cloned downstream to the dually repressed promoter. Fluorescence was measured with different inducer amount (0.1, 0.5, 1.0 and 2 mM IPTG). Promoter activity was found to be maximum after 3 hours of induction by 1mM IPTG.
Input: 1 mM IPTG
Output: Green Fluorescence (501-520 nm)
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Experiment |
Characteristic |
Value |
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Promoter Activity in DH5a Pro cells |
Maximum Fluorescence |
471758.4 (Normalized fluorescence at OD600 ~1.0) 3 hours post induction. |
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Minimum Fluorescence |
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241936.3 (Normalized fluorescence at OD600 ~0.5) 5 minutes post induction
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Promoter Activity in TOP 10 cells |
Maximum Fluorescence |
170546.0 (Normalized fluorescence at OD600 ~1.0) 3 hours after the growth. |
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Growth of culture in LB media |
Doubling time of the recombinant cells |
~ 25 Minutes |
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Response time (after addition of IPTG) |
~20 Minutes |
Compatibility
Chassis: Device has been shown to work in
BBa_E0040, Bba_0010, and Bba_0010
Plasmids: Device has been shown to work on pSB3K3
| None |