Difference between revisions of "Part:BBa K3866038"

Revision as of 09:25, 16 October 2021

J23115:sfGFP:Terminator

Usage and Biology

This TU includes the sfGFP gene placed under the control of the constitutive Anderson promoter J23115 BBa_J23115.

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 a PDGB3_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

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of α1-sbm with: EcoRI, Expected bands: 6345bp, 2947bp, 597bp, Positive result: C2

Experimental Use and Experience

This part showed functionality at the following parts: BBa_K3866029, BBa_K3866031

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 192
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 ===Usage and Biology===
-This TU includes the <i>sfGFP</i> gene placed under the control of the constitutive Anderson promoter J23115 <bbpart>BBa_J23115</bbpart>. Initially, the level 1 cloning was performed using the trc promoter <bbpart>BBa_K3866001</bbpart>, 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 as expected.
+This TU includes the <i>sfGFP</i> gene placed under the control of the constitutive Anderson promoter J23115 <bbpart>BBa_J23115</bbpart>.
 ===Design Notes===