Part:BBa_K1897001:Design
HasA hemophore expression unit
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 480
Design Notes
The presence of a hexa histidine tag would allow for easy protein purification and detection using anti-his antibodies. The promoter BBa_K525998 was chosen as we wished to induce high levels of HasA for purification thus we required a T7 promoter that could be used to induce expression at high levels in E. coli strain BL21(DE3). The terminator BBa_B0015 was chosen as it is commonly used and has a high efficacy.
Source
The HasA coding sequence is originally found in the Serratia marcescens and to construct this part, we used the HasA gene that is from the part BBa K1897000 which was synthesised based on the gene sequence from the National Center for Biotechnology Information (NCBI) database (http://www.ncbi.nlm.nih.gov/nuccore/X81195.2). The Promoter, RBS and terminator are biobrick parts that were obtained from the Distribution kits from iGEM.