Difference between revisions of "Part:BBa K389011:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | * medium strong constitutive promoter, | + | * medium strong constitutive promoter |
| − | * double terminator (forward) to keep | + | * The ''virG'' gene is mutated so it works without an ''rpoA'' subunit from ''Agrobacterium tumefaciens'' (YC Jung ''et al.'', 2004) |
| + | * double terminator (forward) to keep expression of kanamycin resistance by the constitutive promoter low | ||
** double terminator <partinfo>B0017</partinfo> instead of <partinfo>B0015</partinfo> because of problems mentioned with <partinfo>B0012</partinfo> | ** double terminator <partinfo>B0017</partinfo> instead of <partinfo>B0015</partinfo> because of problems mentioned with <partinfo>B0012</partinfo> | ||
| − | * | + | * Kanamycin resistance for screening with agar plates - the more the ''vir'' promoter is activated by VirG the more resistant the cells become to kanamycin. |
===Source=== | ===Source=== | ||
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===References=== | ===References=== | ||
| + | YC Jung ''et al.'' (2004) Mutants of ''Agrobacterium tumefaciens virG'' Gene That Activate Transcription of ''vir'' Promoter in ''Escherichia coli'', ''Current Microbiol'' 49:334-340. | ||
Latest revision as of 14:45, 16 October 2010
VirA screening device
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
- medium strong constitutive promoter
- The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
- double terminator (forward) to keep expression of kanamycin resistance by the constitutive promoter low
- Kanamycin resistance for screening with agar plates - the more the vir promoter is activated by VirG the more resistant the cells become to kanamycin.
Source
- virG gene synthesized by Mr. Gene (BBa_K389002)
- vir promoter from A. tumefaciens C58 (BBa_K389003)
- kanR gene made from BioBrick BBa_P1003
- constitutive promoter and double terminator from parts.igem (BBa_J23110, BBa_B0017)
References
YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.