Device

Part:BBa_K863205:Design

Designed by: Julia Schirmacher   Group: iGEM12_Bielefeld-Germany   (2012-09-23)

GFP integrated in shuttle vector pECPP11JS (BBa_K863204) for integration in yeast


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4772
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4067
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3
    Illegal BglII site found at 2218
    Illegal BamHI site found at 2575
    Illegal XhoI site found at 1198
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4772
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4772
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4928
    Illegal BsaI.rc site found at 1862
    Illegal BsaI.rc site found at 2727


Design Notes

Primers for isolation of GFP were designed with AarI overhanging ends before and after the gene sequence. In the forward primer the start codon has to be deleted.
fwd: 5'-acgtcacctgcgtgtagctCGTAAAGGAGAAGAACTTTT-3'
rev: 5'-AacgtcacctgcgtgtcaagTTATTTGTATAGTTCATCCATG-3'

The PCR product was digested with [http://www.fermentas.de/product_info.php?info=p166&language=de AarI] and ligated in BBa_K863204, which has to be digested with AarI, too. Here you find further information about the design of BBa_K863204 .

Source

This part is composed of GFP (green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP) BBa_E0040 integrated in the shuttle vector pECPP11JS (https://parts.igem.org/Part:BBa_K863204).

References