Difference between revisions of "Part:BBa K4399012"

Revision as of 15:37, 4 October 2022

The inducible anthocyanin biosynthesis system

It is an inducible anthocyanin biosynthesis system, which contains a constitutive GFP expression box, an inducible SbMYB75 expression box, an inducible SbDEL expression box and a constitutive XVE expression box.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1342
Illegal EcoRI site found at 5513
Illegal XbaI site found at 696
Illegal XbaI site found at 1579
Illegal XbaI site found at 8415
Illegal PstI site found at 2759
Illegal PstI site found at 4179
Illegal PstI site found at 4264
Illegal PstI site found at 8599
Illegal PstI site found at 8770
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1342
Illegal EcoRI site found at 5513
Illegal NheI site found at 1244
Illegal PstI site found at 2759
Illegal PstI site found at 4179
Illegal PstI site found at 4264
Illegal PstI site found at 8599
Illegal PstI site found at 8770
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1342
Illegal EcoRI site found at 5513
Illegal BglII site found at 4113
Illegal BglII site found at 4925
Illegal BglII site found at 6994
Illegal BglII site found at 7321
Illegal BglII site found at 8549
Illegal BamHI site found at 3547
Illegal BamHI site found at 3849
Illegal BamHI site found at 5168
Illegal BamHI site found at 5237
Illegal BamHI site found at 8118
Illegal XhoI site found at 7525
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1342
Illegal EcoRI site found at 5513
Illegal XbaI site found at 696
Illegal XbaI site found at 1579
Illegal XbaI site found at 8415
Illegal PstI site found at 2759
Illegal PstI site found at 4179
Illegal PstI site found at 4264
Illegal PstI site found at 8599
Illegal PstI site found at 8770
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1342
Illegal EcoRI site found at 5513
Illegal XbaI site found at 696
Illegal XbaI site found at 1579
Illegal XbaI site found at 8415
Illegal PstI site found at 2759
Illegal PstI site found at 4179
Illegal PstI site found at 4264
Illegal PstI site found at 8599
Illegal PstI site found at 8770
1000
COMPATIBLE WITH RFC[1000]

Results

Construction of level-0 vectors

The DNA elements (golden gate compatible, promoters: P_AtUBI5, P_LexA35S, P_2×35S; CDS of Genes: GFP, SbMYB75, SbDEL, XVE; Terminators: T_mas, T_hsp18.2, T_nos, T_35S) were synthesized by Gensript based on their sequences and were sub-cloned into pUC57.

Construction of level-1 vectors

The level-0 vectors were then used to construct Level-1 vectors: pEC47732: P_AtUBI5-GFP-T_mas, pEC47742: P_LexA35S-SbMYB75- T_hsp18.2, pEC47751: P_LexA35S-SbDEL-T_nos, pEC47761: P_2×35S-XVE-T_35S, according to the protocol:

@@ Line 14: / Line 14: @@
 ===Results===
+== Construction of level-0 vectors ==
 The DNA elements (golden gate compatible, promoters: P<sub>AtUBI5</sub>, P<sub>LexA35S</sub>, P<sub>2×35S</sub>; CDS of Genes: GFP, SbMYB75, SbDEL, XVE; Terminators: T<sub>mas</sub>, T<sub>hsp18.2</sub>, T<sub>nos</sub>, T<sub>35S</sub>) were synthesized by Gensript based on their sequences and were sub-cloned into pUC57.
-The level-0 vectors were then used to contruct Level-1 vectors: pEC47732: P<sub>AtUBI5</sub>- GFP-T<sub>mas</sub>, pEC47742: P<sub>LexA35S</sub>-SbMYB75-T<sub>hsp18.2</sub>, pEC47751: P<sub>LexA35S</sub>-SbDEL-T<sub>nos</sub>, pEC47761: P<sub>2×35S</sub>-XVE-T<sub>35S</sub>, according to the protocol:
+== Construction of level-1 vectors ==
+The level-0 vectors were then used to construct Level-1 vectors: pEC47732: P<sub>AtUBI5</sub>-GFP-T<sub>mas</sub>, pEC47742: P<sub>LexA35S</sub>-SbMYB75- T<sub>hsp18.2</sub>, pEC47751: P<sub>LexA35S</sub>-SbDEL-T<sub>nos</sub>, pEC47761: P<sub>2×35S</sub>-XVE-T<sub>35S</sub>, according to the protocol: