Plasmid

Part:BBa_K4761104

Designed by: Pinhan Pan   Group: iGEM23_HiZJU-China   (2023-09-28)


pCDFDuet dual expression vector

pCDFDuet dual expression vector for rhodioloside production. Vector with a CloDF13 (CDF) origin for the co-expression of two genes. The vector encodes two multiple cloning sites (MCS) each of which is preceded by a T7 promoter, lac operator, and ribosome binding site (rbs).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 112
    Illegal XbaI site found at 2366
    Illegal PstI site found at 131
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 112
    Illegal NheI site found at 1566
    Illegal PstI site found at 131
    Illegal NotI site found at 149
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 112
    Illegal BglII site found at 305
    Illegal BamHI site found at 106
    Illegal XhoI site found at 354
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 112
    Illegal XbaI site found at 2366
    Illegal PstI site found at 131
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 112
    Illegal XbaI site found at 2366
    Illegal PstI site found at 131
    Illegal NgoMIV site found at 324
    Illegal NgoMIV site found at 819
    Illegal AgeI site found at 566
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

pCDFDuet is used for Copy Number Optimization module in 2023 HiZJU-China. The functional parts are linked on the vector using Gibson Assembly. The backbone could be obtained using primers BBa_K4761513 and BBa_K4761514(installing ARO10-adh6) or BBa_K4761518 and BBa_K4761519(installing UGT85A1).

Fig.1 Verification of backbones and genes for Gibson Assembly. pCDFDuet backbone on Lanes 3 & 4.
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