The Gas Vesicle gene cluster (BBa_I750016) submitted by Melbourne 2007 team and the metal sensitive promoter BBa_K190016 were combined, and the product ligated into vector pSB1AC3. The construct was used to test and caracterize the gas vesicle formation.
Melbourne GVP cluster was used containing 400bp repeat in gvpL
This 5.7Kbp part contains 11 open reading frames coding for gas vesicle genes (gvpB,R,N,F,G,L,S,K,J,T and U) from Bacillus megaterium VT1660 AF053765. Original AF053765 DNA in pBluescriptIIKS plasmid (pNL29 in "Gas Vesicle genes identified in Bacillus megaterium and Functional Expression in Escherichia coli, Ning Li and Maura C. Cannon Jounal of bacteriology May 1998 p2450-2458).
This 5.7Kbp part contains 11 open reading frames coding for gas vesicle genes (gvpB,R,N,F,G,L,S,K,J,T and U) from Bacillus megaterium VT1660 AF053765. Promotion of the sequence results in expression of gas vesicles which slow the rate of settling of the cells in Saline.
Site directed mutagenesis was performed in four rounds to remove 3 PstI sites and one EcoRI site from gvpL in the sequence. Primers with biobrick prefix and suffix were then used in a PCR reaction to amplify the part. Finally BBa_J61035 was used to provide the plasmid backbone which introduced a RBS at the front of the part.
10 repeats of 40bp induced durring mutagenesis
Promoter sequence with recognition site for ZntR transcriptional regulator protein.
Conversion of the iGEM parts registry to SBOL2.1