Types | DnaRegion
|
Roles | engineered_region
Composite
|
Sequences | BBa_K571005_sequence (Version 1)
|
Description
The biobrick is composite element, combination of BBa_K571000 and BBa_K571003. The promoter binding to the upstream of acsAB gene, IPTG as the inducer, activates the acsAB gene, producing Cyclic-di-GMP binding domain and UDP-forming domain.
The sequence of acsAB and promoter is amplified by PCR, followed by enzyme digestion and ligation to bind the 2 sequences with the backbone-pSB1C3. The order of restriction enzyme is EcoRI, XbaI and SpeI.
Notes
It is a very long sequence, therefore below is the considerations to take note during cloning.
*The concentration agarose gel during gel electrophoresis should be about 0.8-1%
*The enzyme digestion reaction time is advised to be elongated so that digestion could be more perfect.
*The respective restriction enzyme at the both end of primer should not cut the sequence of another gene
Source
*acsAB: GenBank AB091060.1
*promoter: iGEM registry of standard biological parts