Difference between revisions of "Part:BBa K4201016:Design"

Revision as of 08:54, 12 October 2022

Crte-cytoTDS-MBP_RUBY

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Unknown
12
INCOMPATIBLE WITH RFC[12]
Unknown
21
INCOMPATIBLE WITH RFC[21]
Unknown
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 1002
Illegal PstI site found at 4800
Illegal PstI site found at 5648
Illegal PstI site found at 10856
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 1002
Illegal PstI site found at 4800
Illegal PstI site found at 5648
Illegal PstI site found at 10856
Illegal NgoMIV site found at 7004
Illegal NgoMIV site found at 7583
Illegal NgoMIV site found at 9296
Illegal AgeI site found at 2321
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1
Illegal BsaI site found at 948
Illegal BsaI site found at 5448
Illegal BsaI site found at 5728
Illegal BsaI site found at 6675
Illegal BsaI site found at 10656
Illegal BsaI.rc site found at 934
Illegal BsaI.rc site found at 5434
Illegal BsaI.rc site found at 5714
Illegal BsaI.rc site found at 6661
Illegal BsaI.rc site found at 10642
Illegal BsaI.rc site found at 10922
Illegal SapI.rc site found at 3377
Illegal SapI.rc site found at 3583

Design Notes

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

This construct differs from similar composite parts like 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 assembly was made using NEB 10-beta cells and GoldBio EHA105 agrobacterium as intermediate hosts. Parts BBa_K4201013 and BBa_K4201014 were used as intermediate backbones for Golden Gate assembly. The construct was integrated into part BBa_K4201015 as a final backbone before transfection into Glycine max.

Source

Crte-cytoTDS-MBP is a de novo synthesized construct made for this project. Learn more about our construct and its genes by searching for BBa_K4201004.

RUBY is a reporter gene from the order Caryophyllales.

Gmubi promoters originate from Glycine max.

AtHSP terminators originate from the Arabidopsis thaliana genome.

Synthesis was completed by iGem sponsors IDT and Twist Biosciences.

@@ Line 10: / Line 10: @@
 This construct differs from similar composite parts like BBa_K4201018 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 assembly was made using NEB 10-<i>beta</i> cells and GoldBio EHA105 <i>agrobacterium</i> as intermediate hosts. Parts BBa_K4201013 and BBa_K4201014 were used as intermediate backbones for Golden Gate assembly. The construct was integrated into part BBa_K4201015 as a final backbone before transfection into <i>Glycine max</i>.
 ===Source===