Part:BBa_K197027:Design

Beta Roll

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

This part is in BglBricks Standard. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BglBricks Standard is available at:

[http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BglBricks Standard Description Page]

Source

PCR Odvs0007F/ca1383 on CFT073 gen.           (304bp, gp = A)
PCR ca1382/Odvs0008rR  on CFT073 gen.         (96bp, gp = B)
PCR Odvs0008rF/Odvs0007R on CFT073 gen.       (89 bp, gp = C)
---------------------------------------------------
PCR Odvs0007F/Odvs0007R on A+B+C           (436bp, EcoRI/BamHI)
sub into pBca9495????? .....
---------------------------------------------------
Odvs0007F   Forward EcoRI/BglII for Bglbrick of ECOHLY   ccataGAATTCATGAGATCTtattccgtggaagaacttattgggacc
Odvs0007R   Reverse BamHI for Bglbrick of ECOHLY         ctgatGGATCCataaatatcattaccatatccacc
Odvs0008rF  Removing the BglII site in ECOHLY (Forward)  ggcagtgagggagcagaCctgcttgatggcgg
Odvs0008rR  Removing the BglII site in ECOHLY (Reverse)  ccgccatcaagcagGtctgctccctcactgcc
ca1382	Forward oligo to remove ecoRI site in beta roll	GTTCAGGGcAATTCTCTTGC
ca1383	Reverse oligo to remove ecoRI site in beta roll	GCAAGAGAATTgCCCTGAAC

References

Baumann U, Wu S, Flaherty KM, McKay DB. Three-dimensional structure of the alkaline protease of Pseudomonas aeruginosa: a two-domain protein with a calcium binding parallel beta roll motif. EMBO J. 1993 Sep;12(9):3357-64. PMID: 8253063

Felmlee T, Welch RA. Alterations of amino acid repeats in the Escherichia coli hemolysin affect cytolytic activity and secretion. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5269-73. PMID: 2839840