Plasmid for promoter Amplification & Measurement

RBS +

RBS -

Those plasmids are very useful vectors to amplify promoters and to measure their forces using Standard Promoter Units. The principle is easy to understand. The plasmid contains the BioBrick restriction sites (EcoRI, XbaI, SpeI and PstI) followed by the standard GFP Tripart (E0240). Instead of using a big biobrick containing Promoter, RBS, GFP and Terminators, the Biobrick used for the test is only the Promoter. This plasmid can be used to amplify a promoter because only the promoter is in the biobrick.

One derivated plasmid do not have the RBS B0032 outside the BioBrick site, it allows to test the influence of the RBS in changing it in the BioBrick. To do so, O142 does not contains the RBS, the PCR will begin directly at the ATG codon.

Oligonucleotides design

Creation of 3 oligonucleotides (O140, O141, O142) in order to create two particular plasmids.

Design of the oligonucleotides O140 and O141

Construction of the Plasmid

Necessary material

• Oligos O140, O141 and O142
• E0240
• pSB3K3