DNA

Part:BBa_K1648000

Designed by: Leung King Pong   Group: iGEM15_Hong_Kong-CUHK   (2015-07-07)

Recombination Template for mamAB Operon

Construct Map for K1648000

Consists of four fragments from magnetosome island of Magnetospirillum gryphiswaldense genome.

Due to potential difficulty in introducing the large magnetosome formation operons into Azotobacter vinelandii, a "template" consists of flanking sequences from mamAB operon is designed for homologous recombination.

Part containing this template (K1648002) is introduced into A. vinelandii genome via random integration before full operon sequences to be transformed into A. vinelandii. This Part were verified by double digestion (Fig. 1) and sequencing.

K16480000.jpg

Fig. 1 Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for mamAB Operon (BBa_K1648000) without digestion, with single digestion at XbaI site, with double digestion cut at XbaI and PstI site.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 775
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 291
    Illegal AgeI site found at 810
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 536


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Categories
Parameters
n/aRecombination Template for mamAB Operon