DNA

Part:BBa_K2100061:Design

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


pENTR hEF1a-SV40


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 29
    Illegal EcoRI site found at 1233
    Illegal XbaI site found at 1262
    Illegal PstI site found at 360
    Illegal PstI site found at 865
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 29
    Illegal EcoRI site found at 1233
    Illegal PstI site found at 360
    Illegal PstI site found at 865
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 29
    Illegal EcoRI site found at 1233
    Illegal BglII site found at 614
    Illegal BamHI site found at 1220
    Illegal XhoI site found at 1013
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 29
    Illegal EcoRI site found at 1233
    Illegal XbaI site found at 1262
    Illegal PstI site found at 360
    Illegal PstI site found at 865
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 29
    Illegal EcoRI site found at 1233
    Illegal XbaI site found at 1262
    Illegal PstI site found at 360
    Illegal PstI site found at 865
    Illegal NgoMIV site found at 748
    Illegal AgeI site found at 126
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This basic part entry vector is flanked by L4 and R1 sites, which are used to denote a promoter. This can be cascade with a gene (flanked by L1, L2 sites) using an LR reaction and cloning into a cloned into a backbone that has a negative selection marker between R4 and R2 sites. This part adheres to RFC 65 for recombination based cloning of mammalian parts.


Source

hEF1a comes from human genomic sequence. We ordered hEF1a and SV40 as separate gBlocks from IDT and put them together into a pENTR using Golden Gate assembly.

References