Part:BBa_K3726016
CDS_Lv0_BOH_2_B
This composite part corresponds to a phytobrick compatible MoClo Lv.0 CDS element. This part is an artificial operon designed to express the enzymes CDS_NphT7 (BBa_K3726000), CDS_PhaB^173S (BBa_K3726001), CDS_Ccr (BBa_K3726002), CDS_PhaJ (BBa_K3726003), CDS_PduP (BBa_K3726004), CDS_Slr1192 (BBa_K3726005).
Within this artificial operon each CDS element is preceded by an in-silico designed RBS sequence using DeNovo DNA software. Only the first coding sequence of the operon is not preceded by an RBS sequence which must be assembled during the level 1 MoClo reaction for the expression of the operon in a level 1 transcriptional unit.
This part has been used within the iGEM MADRID_UCM 2021 iGEM team for the expression of a butanol biosynthesis pathway.
In particular, this polycistronic level 0 part codifies for the CDS_PduP and CDS_Slr1192 enzymes of the pathway.
The relative transcription initiation strengths of each RBS in the operon context has been calculated with DeNovo DNA Software. Results are depicted in the graphic below.
For BOH2B
This polycistronic CDS encodes the two last enzymes of the operon whose relative expression level should be lower than the other enzymes. The designed RBS of this operon has been changed from BOH3 part (BBa_K3726011), since expression can now be controlled via the promoter element during the assembly of this part within a transcriptional unit. Then, the RBS sequences have been recalculated to pose a higher strength.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1246
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 252
Illegal NheI site found at 480
Illegal PstI site found at 1246 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1591
Illegal BglII site found at 1708 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1246
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1246
- 1000COMPATIBLE WITH RFC[1000]
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