- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 323
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 61
- 1000Illegal SapI.rc site found at 829
To make the motA biobrick, motA was amplified by PCR using the proofreading Phusion polymerase using DS941 genomic DNA as template. The forward primer incorporated the prefix, added the BBa_B0032 ribosome binding site (RBS) and a scar sequence just upstream of motA and changed the natural GTG start codon to ATG. The reverse primer incorporated the suffix and changed the stop codon to TAA.
Note that an earlier motA biobrick (K777113) started at the first ATG codon within motA and therefore started at the wrong start codon 58 bp into the natural full length motA. Our motA has the correct start as annotated on the E. coli genome sequence and our motA biobrick part BBa_K1463700 is an improvement over K777113.