Generator
Part:BBa_K1598003:Design
Designed by: Marta Napiorkowska Group: iGEM15_UCL (2015-08-27)
RBS-ChAT-6xHis-Terminator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 682
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 303
Illegal XhoI site found at 1770 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1095
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1765
Design Notes
- human CHAT gene (UniProt accession no: P28329) was codon-optimized for E.coli and provided with C-terminal 6xHis tag - due to its length, the sequence was synthesized as two gBlock fragments with 30 bp overlap between each other and with 20 bp overlaps with respective PSB1C3 ends - the construct was cloned into PSB1C3 using Gibson Assembly
Source
DNA synthesis