Difference between revisions of "Part:BBa K4179019"

Revision as of 10:02, 8 October 2022

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_K4179016) as an RBS while XimE has (BBa_K4179018). 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

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1772
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1772
Illegal NheI site found at 1502
Illegal NheI site found at 1995
21
INCOMPATIBLE 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
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1772
25
INCOMPATIBLE 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
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4179019 short</partinfo>
-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_K4179016) as an RBS while XimE has (BBa_K4179018). 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).
+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_K4179016) as an RBS while XimE has (BBa_K4179018). 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===