Plasmid_Backbone

Part:BBa_K4216045

Designed by: Yves Loiseau--Marchand   Group: iGEM22_Paris_Bettencourt   (2022-10-11)


pCOLADuet_Bettencourt

The pDuet_Bettencourt Vectors is a sets of 3 compatible vector for transformation in E.Coli. We re-engineered the pETDuet System (https://www.merckmillipore.com/FR/fr/product/pETDuet-1-DNA-Novagen,EMD_BIO-71146#documentation) and cloned the Transcriptional unit of a GFP from the V35 vector of the CIDAR MoClo[1,2] flanked with BsaI restriction site and barcode A and E of the Moclo for assembly of entire transcriptional unit. Like so, during a Golden Gate Cloning, after the BsaI cut, the GFP is replaced by the insert. This allow for easy selection against Green fluorescence (compatible with the mScarlet reporter used in all of our toolkit)

PCOLA.png

Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 1888
    Illegal SpeI site found at 2120
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NheI site found at 1238
    Illegal NheI site found at 2145
    Illegal NheI site found at 2168
    Illegal SpeI site found at 2120
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 1888
    Illegal SpeI site found at 2120
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 1888
    Illegal SpeI site found at 2120
    Illegal AgeI site found at 150
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 223
    Illegal BsaI site found at 3136
    Illegal BsaI.rc site found at 2101


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