Difference between revisions of "Part:BBa K4201017:Design"

(→‎Design Notes)
(→‎Design Notes)
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This notably does not contain the sequence encoding Maltose-binding protein (MBP), which was done purposely to determine if MBP had an impact on taxadiene production in <i>Glycine max</i>
 
This notably does not contain the sequence encoding Maltose-binding protein (MBP), which was done purposely to determine if MBP had an impact on taxadiene production in <i>Glycine max</i>
 +
 +
This assembly was made using NEB 10-<i>beta</i> cells and GoldBio EHA105 <i>agrobacterium</i> as intermediate hosts. Parts BBa_K4201013 (Amp BsaI) and BBa_K4201014 (Chlor AaRI) were used as intermediate backbones for Golden Gate assembly. The construct was integrated into part BBa_K4201015 (Kan BsaI) as a final backbone before transfection into <i>Glycine max</i>.
  
 
===Source===
 
===Source===

Revision as of 08:57, 12 October 2022


CrtE_cytoTDS2_RUBY


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2087
    Illegal PstI site found at 5638
    Illegal PstI site found at 10846
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2087
    Illegal PstI site found at 5638
    Illegal PstI site found at 10846
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2087
    Illegal PstI site found at 5638
    Illegal PstI site found at 10846
    Illegal NgoMIV site found at 1697
    Illegal NgoMIV site found at 6994
    Illegal NgoMIV site found at 7573
    Illegal NgoMIV site found at 9286
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI site found at 948
    Illegal BsaI site found at 1887
    Illegal BsaI site found at 2167
    Illegal BsaI site found at 3113
    Illegal BsaI site found at 5438
    Illegal BsaI site found at 5718
    Illegal BsaI.rc site found at 934
    Illegal BsaI.rc site found at 1873
    Illegal BsaI.rc site found at 2153
    Illegal BsaI.rc site found at 3100
    Illegal BsaI.rc site found at 5424
    Illegal BsaI.rc site found at 5704
    Illegal BsaI.rc site found at 6651


Design Notes

Part information and design consideration can be found on respective basic parts pages.

This construct differs from similar composite parts like BBa_K4201016 and BBa_K4201018 by having different promoter (Gmubi) and terminator (AtHSP) amounts and locations. In these constructs, dashes(-) represent locations where genes were synthesized de novo. An underscore (_) symbolizes a location where we adhered genes together via Golden Gate assembly. Utilizing Golden Gate assembly allowed us to insert promoter and terminator sequences between genes of interest, and consequently enables the study of transcription efficacy based on promoter and terminator location.

This notably does not contain the sequence encoding Maltose-binding protein (MBP), which was done purposely to determine if MBP had an impact on taxadiene production in Glycine max

This assembly was made using NEB 10-beta cells and GoldBio EHA105 agrobacterium as intermediate hosts. Parts BBa_K4201013 (Amp BsaI) and BBa_K4201014 (Chlor AaRI) were used as intermediate backbones for Golden Gate assembly. The construct was integrated into part BBa_K4201015 (Kan BsaI) as a final backbone before transfection into Glycine max.

Source

Gmubi promoters originate from Glycine max.

AtHSP terminators originate from the Arabidopsis thaliana genome.

CrtE is a GGPP synthase from Pantoea ananatis LMG 20103

cytoTDS2 is a taxadiene synthase native to Taxus chinensis var. mairei optimized for use in Glycine max.

RUBY is a reporter gene from the order Caryophyllales.