Part:BBa_K4179019
XimD + XimE under T7 promoters + LacI + Lac operator
This composite part comprises of flavin-dependent monooxygenase - XimD (BBa_K4179016) and SnoaL-like cyclase - XimE (BBa_K4179018), under the T7 promoter (BBa_J64997). XimD has (BBa_K2906017) as an RBS while XimE has (BBa_K3875029). XimE is constituvely expressed. Between the promoter and RBS of XimD there is a Lac operator (BBa_K1624002) that makes the expression of XimD to be under the regulation of LacI (BBa_C0012). LacI is cloned under the native pLacI promoter (BBa_K3842018).
Use and purpose
The purpose of the composite part is to introduce XimD and XimE into E. coli to allow the formation of decursinol (a pyran) from 7-methyldesuberosin. To ensure more of the pyran product (decursinol) in comparison with the furan product (marmesin), the team constrained a surplus of XimE via genetic manipulations.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1772
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1772
Illegal NheI site found at 1502
Illegal NheI site found at 1995 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1772
Illegal BglII site found at 3282
Illegal BamHI site found at 1457
Illegal XhoI site found at 1189 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1772
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1772
Illegal NgoMIV site found at 178
Illegal NgoMIV site found at 1237
Illegal AgeI site found at 1751 - 1000COMPATIBLE WITH RFC[1000]
None |