Difference between revisions of "Part:BBa K3866026"

Revision as of 22:10, 25 September 2021

ParaBAD:sbm:Terminator

Usage and Biology

This TU includes the sbm gene placed under the control of the arabinosed-induced promoter. Initially, the level 1 cloning was performed using the trc promoter, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working more than well.

Design Notes

The coding sequence was domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in seva α1 vector and has overhangs compatible for GoldenBraid cloning.

Figure 1. The first level α module of the Propionate Production: α1:ParaBAD:RBS-smb-Double terminator

Verification of Cloning

Fig.2:: (U=Uncut , C= Cut)Positive clone: 6,Seva a2 arac ACK. Expected bands 4601, 853

Experimental Use and Experience

This part showed functionality at this part BBa_K3866009

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 2948
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 2948
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 2948
Illegal BamHI site found at 1148
Illegal BamHI site found at 2236
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 2948
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 2948
Illegal NgoMIV site found at 3263
Illegal AgeI site found at 983
Illegal AgeI site found at 1400
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 965

@@ Line 4: / Line 4: @@
 ===Usage and Biology===
-This TU includes the AckA gene placed under the control of the arabinosed-induced promoter. Initially, the level 1 cloning was performed using the trc promoter, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working more than well.
+This TU includes the <i>sbm</i> gene placed under the control of the arabinosed-induced promoter. Initially, the level 1 cloning was performed using the trc promoter, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working more than well.
 ===Design Notes===