Difference between revisions of "Part:BBa K814001"

Latest revision as of 14:28, 9 August 2020

ATP-grasp (ATPG) generator

Our research has focused on two novel biosynthetic pathways found in two distinct algal species. A pathway ending in the production of two UV-protective compounds, shinorine and mycosporine-glycine, was cloned from Anabaena varibalis. ATP-grasp (ATPG) catalyzes the third step in this pathway, 4-deoxygadusol into mycosporine-glycine.

This part includes a modified constitutive lac promoter (lacP'), and RBS and the open reading frame of DHQS. This part can be used to express DHQS in E. coli.

Figure 1. PCR screen of ATP-grasp confirming successful cloning of this ORF into the pUCBB plasmid.

Balskus and Walsh, 2010. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3116657/]

BBa_K3634002

Codon optimised adenosine triphosphate (ATP)-grasp (ATPG) CDS for use in E.coli as part of the shinorine synthesis gene cluster. By using the IDT codon optimisation tool, St Andrews iGEM 2020 have optimised the GC content of the sequence taken from A.variabilis to improve production efficiency of the final product shinorine. We have further made the part biobrick assembly standard RFC[10] & RFC[1000] compatible by removing EcoR1 and PstI restriction sites introduced by this optimisation step. The part should be used alongside the additional optimised parts (BBa_K3634000, BBa_K3634001 and BBa_K3634003) responsible for the ultimate conversion of the substrate sedoheptulose 7-phosphate to the final product shinorine.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 944
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 944
Illegal NotI site found at 1517
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 944
Illegal BglII site found at 134
Illegal XhoI site found at 1525
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 944
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 944
1000
COMPATIBLE WITH RFC[1000]

@@ Line 11: / Line 11: @@
 Balskus and Walsh, 2010. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3116657/]
+== BBa_K3634002 ==
+Codon optimised adenosine triphosphate (ATP)-grasp (ATPG) CDS for use in E.coli as part of the shinorine synthesis gene cluster. By using the IDT codon optimisation tool, St Andrews iGEM 2020 have optimised the GC content of the sequence taken from A.variabilis to improve production efficiency of the final product shinorine. We have further made the part biobrick assembly standard RFC[10] & RFC[1000] compatible by removing EcoR1 and PstI restriction sites introduced by this optimisation step. The part should be used alongside the additional optimised parts (BBa_K3634000, BBa_K3634001 and BBa_K3634003) responsible for the ultimate conversion of the substrate sedoheptulose 7-phosphate to the final product shinorine.
 <!-- Add more about the biology of this part here