Difference between revisions of "Part:BBa K3279001"
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===Usage and Biology=== | ===Usage and Biology=== | ||
+ | Firstly, the CrtB gene was amplified from Rhodobacter sphaeroides's genomic sequence by PCR (see Figure 1a) and was cloned into pET-30a(+) plasmid. Then, colony PCR was run to check the success of ligation (see Figure 1b). | ||
+ | [[File:CAU CrtB Fig1.png|300px|thumb|center|'''Fig. 1''' pET-CrtB plasmid construction. (a) Amplified CrtB from Rhodobacter sphaeroides's genomic sequence; (b) Colony PCR verified that CrtB was successfully assembled into the pET-30a (+) vector.]] | ||
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Revision as of 04:20, 19 October 2019
Phytoene Synthase (CrtB) from Rhodobacter sphaeroides
This part is a phytoene synthase which transforms geranylgeranyl diphosphate into 15-cis-phytoene. It is a crucial enzyme in lycopene synthesis pathway and always works together with geranylgeranyl diphosphate synthase (CrtE) and phytoene desaturase (CrtI). This sequence is optimized for use in E.coli.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 245
Illegal NotI site found at 968 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1025
Illegal XhoI site found at 334
Illegal XhoI site found at 1006 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 264
Illegal NgoMIV site found at 722 - 1000COMPATIBLE WITH RFC[1000]