Difference between revisions of "Part:BBa K3866030"

Revision as of 23:26, 25 September 2021

ParaBAD:ygfG:Terminator - ParaBAD:ygfH:Terminator

Usage and Biology

This composite part consists of the following Transcription Units:

ygfG: BBa_K3866006 & ygfH: BBa_K3866019

Design Notes

The coding sequences were domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequences are cloned in seva ω2 vector and have overhangs compatible for GoldenBraid cloning.

Figure 1. The ω module of the Propionate Production: ω2:ParaBAD:RBS-ygfG-Double terminator:ParaBAD:RBS-ygfH-Double terminator

Verification of Cloning

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of ω1-sbm-stuffer (C6-C7-C8) with: EcoRI + EcoRV, Expected bands: 7439bp, 1509bp, 1132bp, 302bp Positive result: C6-C7-C8

Experimental Use and Experience

This part showed functionality at part BBa_K3866031

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 4655
Illegal BamHI site found at 1148
Illegal BamHI site found at 1424
Illegal BamHI site found at 3343
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 983
Illegal AgeI site found at 3178
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 965
Illegal SapI site found at 3160
Illegal SapI.rc site found at 1883

References

Akawi L, Srirangan K, Liu X, Moo-Young M, Perry Chou C. Engineering Escherichia coli for high-level production of propionate. J Ind Microbiol Biotechnol. 2015 Jul;42(7):1057-72. https://doi.org/10.1007/s10295-015-1627-4

@@ Line 6: / Line 6: @@
 This composite part consists of the following Transcription Units:
-<p><i>ygfG</i> <bbpart>BBa_K3866006</bbpart> and <i>ygfH</i> <bbpart>BBa_K3866019</bbpart>
+<p><i>ygfG</i>: <bbpart>BBa_K3866006</bbpart> & <i>ygfH</i>: <bbpart>BBa_K3866019</bbpart>
 ===Design Notes===