DNA

Part:BBa_K2100071

Designed by: Colleen Foley   Group: iGEM16_MIT   (2016-10-19)


pEXPR hEF1a: attB-flipped eYFP-attP

This expression vector contains pENTR hEF1a, constitutively expressed mammalian promoter human elongation factor 1 alpha, and pENTR attB-flipped eYFP-attP, an entry vector containing attB and attP recognition sites for recombinase TP901, with an inverted yellow fluorescent protein, eYFP, between them.

The flipped eYFP gene is oriented upside down, preventing translation of the gene. The flipped fluorescent gene is flanked by recognition sites of TP901 recombinase, which is oriented in a way that only allows unidirectional flip. When recombinase TP901 is present, they will bind to the recognition sites and flip the upside-down eYFP to the correct orientation, allowing the fluorescent gene to be expressed.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal EcoRI site found at 3389
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal PstI site found at 3098
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal EcoRI site found at 3389
    Illegal NheI site found at 2071
    Illegal NheI site found at 2337
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal PstI site found at 3098
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal EcoRI site found at 3389
    Illegal BglII site found at 591
    Illegal BamHI site found at 1197
    Illegal BamHI site found at 2501
    Illegal XhoI site found at 990
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal EcoRI site found at 3389
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal PstI site found at 3098
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 1210
    Illegal EcoRI site found at 3389
    Illegal PstI site found at 337
    Illegal PstI site found at 842
    Illegal PstI site found at 3098
    Illegal NgoMIV site found at 725
    Illegal AgeI site found at 103
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1948


[edit]
Categories
Parameters
None