Coding

Part:BBa_K3279001

Designed by: Shujie Liao   Group: iGEM19_CAU_China   (2019-10-13)
Revision as of 05:15, 19 October 2019 by JIAO (Talk | contribs) (→‎Usage and Biology)


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.

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).

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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 245
    Illegal NotI site found at 968
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1025
    Illegal XhoI site found at 334
    Illegal XhoI site found at 1006
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 264
    Illegal NgoMIV site found at 722
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None