Part:BBa_K5299203

BBa_J23119 - BBa_B0034 - BBa_I746916 - BBa_B0015

Description

It consists of a promoter, a RBS, a sfGFP and a terminator. The BBa_J23119 is a constitutive Anderson promoter.

Usage and Biology

It is able to produce the sfGFP according to the promoter's abilities.

It was constructed in order to check the strength of the P3.1 promoter. For our experiments, this construct was used as a phase activation control for the P3.1 promoter ( BBa_K4583008. )

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 61
  

Results

Measurements for OD 600nm and fluorescence (488nm,515nm) were taken over the course of 16 hours, in a 96-well microplate. Clonings were done according to the Golden Braid method, leaving us with level α constructs in the pDGB3a1 backbone BBa_K4213058.

Constructs and pDGB3a1 (empty vector) transformed into E.coli BL21 (DE3) chassis, incubated at 37oC, 180rpm for 16 hours.

Plated 200 ul 5 times, out of each single liquid bacterial culture, created from the same bacterial colony, in order to establish accuracy through technical repeats.

Medium used was M9 due to minimal interference, with D-glucose serving as the carbon source. Also, served as blank, plated 5 times.

Measurements were normalised as such: using the average price of fluorescence for the 5 technical repeats and dividing it by the average price of OD.
Standard deviation included in the graphs.

Measurements were taken over the course of 16 hours. Here are the results for this part (mentioned in the graph as J23119 B0034).

Figure 1: Constructs with the BBa_J23119 or the P3.1 (BBa_K4583008) or the BBa_J45992 promoter, the BBa_B0034 RBS, the BBa_I746916 sfGFP and the BBa_B0015 terminator. pDGB3a1 serves as a negative control. BBa_J23119 and BBa_J45992 serve as phase activation controls. BBa_J45992 also serves as strength control, due to the same phase activation with BBa_K4583008.