Plasmid

Part:BBa_K4213013

Designed by: Ioannis Retalis   Group: iGEM22_Thessaly   (2022-08-27)


pENTR-GFP-aHHR_theophylline

The GFP fluorescent reporter protein under the regulation of a synthetic theophylline aptazyme, in the pENTR plasmid vector. We used the vector to "extract" the aptazyme sequence with PCR and use it in our experiments. We used four primers because there was a BsmbI restriction site inside the sequence and we had to domesticate the part. We retrieved the aptazyme with the terminator and created a Golden Braid B6-C1 part.

You can find the primers we used here: ...

Disregard the Assembly Compatibility notification, because this is a whole plasmid. Domesticate each of its parts depending on the cloning method that you want to use.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1496
    Illegal XbaI site found at 1670
    Illegal SpeI site found at 2412
    Illegal PstI site found at 2390
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1496
    Illegal NheI site found at 239
    Illegal NheI site found at 505
    Illegal NheI site found at 721
    Illegal NheI site found at 2404
    Illegal SpeI site found at 2412
    Illegal PstI site found at 2390
    Illegal NotI site found at 671
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1496
    Illegal BamHI site found at 1650
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1496
    Illegal XbaI site found at 1670
    Illegal SpeI site found at 2412
    Illegal PstI site found at 2390
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1496
    Illegal XbaI site found at 1670
    Illegal SpeI site found at 2412
    Illegal PstI site found at 2390
    Illegal NgoMIV site found at 2141
    Illegal AgeI site found at 730
    Illegal AgeI site found at 2014
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 116


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