Not Released
Sample In stock
Experience: Works
Not Used
Get This Part
Composite

Part:BBa_K1632013:Design

Designed by: Riku Shinohara   Group: iGEM15_Tokyo_Tech   (2015-08-30)
Revision as of 13:49, 15 September 2015 by JunKawamura (Talk | contribs) (→‎Materials and Methods)

PBAD/araC_rbs_fimE(wild-type)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1260
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961

Design Notes

sequence confirmed

Materials and Methods

1. Construction
All the samples were DH5α strain with antibiotic resistance to ampicillin and kanamycin.

A. PBAD/araC_fimE(wild-type) (6A1) +fim switch[default ON](wild-type)_rbs_gfp (3K3)
B. PBAD/araC_fimE(wild-type) (6A1) +fim switch[default ON](wild-type)_rbs_gfp (3K3)
C. rbs_M256IcysE (6A1) + fim switch[default ON](wild-type)_rbs_gfp (3K3) …positive control 1
D. rbs_M256IcysE (6A1) + fim switch[default OFF](wild-type)_rbs_gfp (3K3) …negative control 1
E. PBAD/araC_fimE(wild-type) (6A1) + J23119_rbs_gfp (3K3) …positive control 2
F. PBAD/araC_fimE(wild-type) (6A1)+rbs_gfp (3K3) …negative control 2

Fig. 1. Plasmids

2. Assay protocol

Source

Composite of BBa_I0500, BBa_B0034, BBa_K1632011

References

Timothy S. Ham et al. (2006) A Tightly Regulated Inducible Expression System Utilizing the fim Inversion Recombination Switch. Biotechnol Bioeng 94(1):1-4