Difference between revisions of "Part:BBa K4201018:Design"

(→‎Design Notes)
(→‎Design Notes)
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This construct differs from similar composite parts like BBa_K4201016 by having different promoter (Gmubi) and terminator (AtHSP) amounts and locations. In these constructs, dashes(-) represent locations where genes were synthesized <i>de novo</i>. 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 construct differs from similar composite parts like BBa_K4201016 by having different promoter (Gmubi) and terminator (AtHSP) amounts and locations. In these constructs, dashes(-) represent locations where genes were synthesized <i>de novo</i>. 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.
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RUBY is a reporter gene that enables the visualization of transformation by changing the color of the plant from green to purle-red<sup>4</sup>.
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Gmubi is constitutive promoter native to <i>Glyine max</i><sup>5</sup>, while AtHSP is the terminator of a heat shock protein that has shown to promote expression in plants<sup>6</sup>.
  
 
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>.
 
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>.

Revision as of 10:20, 12 October 2022


CrtE_cytoTDS2-MBP_RUBY


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4087
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4087
    Illegal PstI site found at 2087
    Illegal PstI site found at 3774
    Illegal PstI site found at 5335
    Illegal PstI site found at 6893
    Illegal PstI site found at 12101
    Illegal NgoMIV site found at 1697
    Illegal NgoMIV site found at 8249
    Illegal NgoMIV site found at 8828
    Illegal NgoMIV site found at 10541
    Illegal AgeI site found at 3923
  • 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 3114
    Illegal BsaI site found at 6693
    Illegal BsaI site found at 6973
    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 6679
    Illegal BsaI.rc site found at 6959
    Illegal BsaI.rc site found at 7906


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 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.

RUBY is a reporter gene that enables the visualization of transformation by changing the color of the plant from green to purle-red4.

Gmubi is constitutive promoter native to Glyine max5, while AtHSP is the terminator of a heat shock protein that has shown to promote expression in plants6.

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

cytoTDS-MBP is a de novo synthesized construct made for this project. Learn more about our construct and its genes by searching for BBa_K4201006.

RUBY is a reporter gene from the order Caryophyllales.

Gmubi promoters originate from Glycine max.

AtHSP terminators originate from the Arabidopsis thaliana genome.

All genes were synthesized de novo by iGEM sponsors Twist Biosciences and IDT.