Generator

Part:BBa_I750016:Design

Designed by: Phillip Dodson   Group: iGEM07_Melbourne   (2007-10-21)
Revision as of 12:46, 21 October 2007 by PhillipDodson (Talk | contribs) (→‎Source)


Gas Vesicle polycistonic gene


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 334
    Illegal BamHI site found at 4139
    Illegal XhoI site found at 3827
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 785
    Illegal NgoMIV site found at 1187
    Illegal AgeI site found at 4106
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2597
    Illegal SapI.rc site found at 3753


Design Notes

Site directed mutagenesis was performed in four rounds to remove 3 PstI sites and one EcoRI site from gvpL in the sequence. Primers with biobrick prefix and suffix were then used in a PCR reaction to amplify the part. Finally BBa_J61035 was used to provide the plasmid backbone which introduced a RBS at the front of the part.


Source

This 5.7Kbp part contains 11 open reading frames coding for gas vesicle genes (gvpB,R,N,F,G,L,S,K,J,T and U) from Bacillus megaterium VT1660 AF053765. The original DNA (sequence= AF053765) was provided by Maura Cannon in pBluescriptIIKS plasmid (pNL29 in "Gas Vesicle genes identified in Bacillus megaterium and Functional Expression in Escherichia coli", Ning Li and Maura C. Cannon, Jounal of bacteriology, May 1998 p2450-2458). Site dirrected mutagenesis was performed to produce four silent changes, removing restriction sites.

References

pNL29 in "Gas Vesicle genes identified in Bacillus megaterium and Functional Expression in Escherichia coli", Ning Li and Maura C. Cannon, Jounal of bacteriology, May 1998, p2450-2458