Difference between revisions of "Part:BBa K1632013:Design"

(→‎Materials and Methods)
(→‎Materials and Methods)
Line 10: Line 10:
 
All the samples were DH5α strain with antibiotic resistance to ampicillin and kanamycin.<br>
 
All the samples were DH5α strain with antibiotic resistance to ampicillin and kanamycin.<br>
  
−
A. PBAD/araC_FimE(wild-type) (6A1) +fim switch[default ON](wild-type)_gfp (3K3) <br>
+
A. PBAD/araC_fimE(wild-type) (6A1) +''fim'' switch[default ON](wild-type)_rbs_gfp (3K3) <br>
−
B. PBAD/araC_FimE(wild-type) (6A1) +fim switch[default ON](wild-type)_gfp (3K3) <br>
+
B. PBAD/araC_fimE(wild-type) (6A1) +''fim'' switch[default ON](wild-type)_rbs_gfp (3K3) <br>
−
C. rbs_M256IcysE (6A1) + fim switch[default ON](wild-type)_gfp (3K3) …positive control 1<br>
+
C. rbs_M256IcysE (6A1) + ''fim'' switch[default ON](wild-type)_rbs_gfp (3K3) …positive control 1<br>
−
D. rbs_M256IcysE (6A1) + fim switch[default OFF](wild-type)_gfp (3K3) …negative control 1<br>
+
D. rbs_M256IcysE (6A1) + ''fim'' switch[default OFF](wild-type)_rbs_gfp (3K3) …negative control 1<br>
−
E. PBAD/araC_FimE(wild-type) (6A1) + J23119_gfp (3K3) …positive control 2 <br>
+
E. PBAD/araC_fimE(wild-type) (6A1) + J23119_rbs_gfp (3K3) …positive control 2 <br>
−
F. PBAD/araC_FimE(wild-type) (6A1)+rbs_gfp (3K3) …negative control 2 <br>
+
F. PBAD/araC_fimE(wild-type) (6A1)+rbs_gfp (3K3) …negative control 2 <br>
 
[[Image:Tokyo_Tech_FimE_assay.png|thumb|center|600px|<b>Fig. 1. </b>Plasmids]]<br>
 
[[Image:Tokyo_Tech_FimE_assay.png|thumb|center|600px|<b>Fig. 1. </b>Plasmids]]<br>
  

Revision as of 13:49, 15 September 2015

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