Composite

Part:BBa_K863207:Design

Designed by: Isabel Huber   Group: iGEM12_Bielefeld-Germany   (2012-09-26)
Revision as of 13:06, 26 September 2012 by Isahu (Talk | contribs) (→‎Design Notes)

tvel5 integrated in shuttle vector pECPP11JS (BBa_K863204) for integration in yeast


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 5558
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4853
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3
    Illegal BglII site found at 1931
    Illegal BglII site found at 2363
    Illegal BglII site found at 3004
    Illegal BamHI site found at 3361
    Illegal XhoI site found at 1198
    Illegal XhoI site found at 1963
    Illegal XhoI site found at 2596
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 5558
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 5558
    Illegal NgoMIV site found at 2449
    Illegal NgoMIV site found at 2602
    Illegal AgeI site found at 1330
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5714
    Illegal BsaI.rc site found at 1885
    Illegal BsaI.rc site found at 3513
    Illegal SapI.rc site found at 1395


Design Notes

Design Notes

Primers for isolation of tvel5 were designed with AarI overhanging ends before and after the gene sequence.
fwd: 5'-acgtcacctgcgtgtagctggtatcggtcctgtcgccg-3'
rev: 5'-acgtcacctgcgtgtcaagTTACTGGTCGCTCGGGTCGC-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. For further information about the design of BBa_K863204 look here.

Source

coming soon

References