Composite

Part:BBa_K3365017:Design

Designed by: JUNYAN LI   Group: iGEM20_SJTU-BioX-Shanghai   (2020-10-23)
Revision as of 15:44, 27 October 2020 by Little ghost (Talk | contribs) (→‎References)

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pBAD upstream of eGFP with inhibition unit containing lure3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1053
    Illegal XhoI site found at 1062
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 74
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 56


Design Notes

The restriction cutting sites are added to the fragment during PCR for easy ligation to the backbone.

Source

The ligation of BBa_K3365003, BBa_K3365054 and BBa_K3365002 by Overlap PCR.

References

[1] Guzman LM, Belin D, Carson MJ, Beckwith J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol. 1995 Jul;177(14):4121-30. [2] Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA. Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell. 2013 Feb 28;152(5):1173-83. [3] Vigouroux A, Bikard D. CRISPR Tools To Control Gene Expression in Bacteria. Microbiol Mol Biol Rev. 2020 Apr 1;84(2):e00077-19.