Difference between revisions of "Part:BBa K2205026"

Revision as of 19:21, 27 October 2017

pSB1A51 - Promoter Library Screening Plasmid

Screening plasmid for the construction and screening of a biobrick compatible promoter library. This part is an improvement of a previous promoter library screening plasmid (J61002). The improvements made are as follows:

1 – A bi-directional terminator has been inserted upstream of the iGEM cloning site to prevent any leaky expression from the plasmid backbone and to prevent expression from the promoter in the reverse direction.

2 – The Lac Operator sequence has been inserted immediately downstream of the promoter insertion site, this is to allow for the expression of the promoter to be repressed until the user desires to test the construct.

3 – The ribosome binding site (B0034) is inserted between two restriction sites (SacI and BamHI). This is to allow for the RBS to be changed at will if the user wishes to screen a library of RBS as well as promoters.

4 – The reporter gene used is sfGFP, this is a shorter gene sequence which produces a brighter fluorescence than mRFP. The emission wavelength of sfGFP is (509 nm) which is further from the standard cell density wavelength (600 nm) than the emission wavelength of mRFP (584 nm), this makes interference from high cell density samples less likely.

Sequence and Features