Device

Part:BBa_K2873003:Design

Designed by: Zhipeng Ding   Group: iGEM18_Minnesota   (2018-10-10)


LacI-glnA-GFP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1375
    Illegal EcoRI site found at 1645
    Illegal PstI site found at 2666
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1375
    Illegal EcoRI site found at 1645
    Illegal PstI site found at 2666
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1375
    Illegal EcoRI site found at 1645
    Illegal BglII site found at 1961
    Illegal BamHI site found at 2337
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1375
    Illegal EcoRI site found at 1645
    Illegal PstI site found at 2666
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1375
    Illegal EcoRI site found at 1645
    Illegal PstI site found at 2666
    Illegal AgeI site found at 1803
    Illegal AgeI site found at 2238
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3329


Design Notes

LacI-glnA-GFP)is constructed from pSB1C3 plasmid backbone consisting LacI protein coding region, LacI binding site, glnA coding region and GFP. LacI protein coding region overexpression of LacI to ensure inhibition of LacI promoter LacI binding site: controlled promoter region that only operates when LacI is released from IPTG binding glnA coding region: expression of glutamine synthetase GFP: Green fluorescent protein marker for characterizing the expression level of glnA

Source

Gibson assembly product sourced from existing plasmids in Zhang lab.

References