This is a list of all BioBricks we used from the registry or created by ourselves and a short explanation, why we used them. The list is sorted alphabetically.
| BioBrick (name and partnumber)
| Status
| Comment
|
| double terminator, <partinfo>B0017</partinfo>
| from registry
|
- this contains two <partinfo>B0010</partinfo> terminators
- we used this because of problems mentioned in registry with <partinfo>B0012</partinfo>
|
| RBS, <partinfo>B0034</partinfo>
| from registry
|
- we always assembled a strong RBS
|
| lacI gene, <partinfo>C0012</partinfo>
| from registry
|
- used this to build the tightly regulated lac-operon <partinfo>K389050</partinfo>
|
| mRFP, <partinfo>E1010</partinfo>
| from registry
|
- this BioBrick was used as a reporter gene, e.g. in the construct <partinfo>K389013</partinfo>
|
| lacIq promoter, <partinfo>I14032</partinfo>
| from registry
|
- used this to build the tightly regulated lac-operon <partinfo>K389050</partinfo>
|
| mRFP generator, <partinfo>J04450</partinfo>
| from registry
|
- this BioBrick was used as a visible selection marker for cloning other BioBricks into the <partinfo>pSB1C3</partinfo> plasmid
|
| constitutive promoter, <partinfo>J23110</partinfo>
| from registry
|
- we always used a medium strong constitutive promoter from the registry to express our BioBricks
- this BioBrick was used e.g. in <partinfo>K389010</partinfo> or <partinfo>K389011</partinfo>
|
| R6K origin, <partinfo>J61001</partinfo>
| from registry
|
- this BioBrick was used to create a plasmid without ColE1 ori
- the R6K ori only works in some E. coli strains, so it is used for a two plasmid screening system
|
| virA, <partinfo>K238008</partinfo>
| from registry, fixed
|
- we wanted to use the virA BioBrick from the registry but problems occurred, so we created our own virA from Agrobacterium tumefaciens C58 TI-plasmid
- our own (working) virA is here: <partinfo>K389001</partinfo>
|
| vir-promoter, <partinfo>K238011</partinfo>
| from registry, fixed
|
- same as the virA BioBrick - this one from the registry does not work properly, so we made a vir-promoter by ourselves
- the part that was sent to us is actually a tetR gene (<partinfo>C0040</partinfo>) under the control of a constitutive promoter (<partinfo>J23105</partinfo>) before a double terminator (<partinfo>B0015</partinfo>, compare our sequencing results)
|
| virA, <partinfo>K389001</partinfo>
| new
|
- "our" virA
- illegal restriction site removed by site-directed mutagenesis
- isolated from A. tumefaciens C58 TI-plasmid
|
| virG, <partinfo>K389002</partinfo>
| synthesized, new
|
- virG that works in E. coli without rpoA gene from A. tumefaciens
- ...
|
| vir-promoter, <partinfo>K389003</partinfo>
| new
|
- "our" vir-promoter
- isolated from A. tumefaciens C58 TI-plasmid
|
| firefly luciferase, <partinfo>K389004</partinfo>
| new
|
- "our" luciferase
- sensitive reporter gene
- isolated from Promega's pGL4.10luc2 plasmid
|
| Kanamycin resistance gene, <partinfo>K389005</partinfo>
| new
|
- "our" antibiotic resistance
- used for directed evolution and mutated virA screenings, respectively
- isolated from the BioBrick <partinfo>P1003</partinfo>
|
| virA generator, <partinfo>K389010</partinfo>
| composite
|
- medium strong constitutive promoter, so there is enough VirA receptor expressed but not too much so the cell suffers from the expression
- ...
|
| virA screening device, <partinfo>K389011</partinfo>
| composite
|
- the new virG which works without rpoA and a kanamycin resistance gene under the control of a vir promoter to screen mutants of virA (which are on a different plasmid in a <partinfo>K389010</partinfo>-like part)
- the better the VirA receptor recognizes a substance the more resistant the cell is against kanamyin
|
| virA reporter device with luc, <partinfo>K389012</partinfo>
| composite
|
|
| virA reporter device with mRFP, <partinfo>K389013</partinfo>
| composite
|
|
| Test, <partinfo>K389014</partinfo>
| composite
|
|
| vir promoter characterisation part with luc, <partinfo>K389015</partinfo>
| composite
|
|
| vir promoter characterisation part with mRFP, <partinfo>K389016</partinfo>
| composite
|
| vir promoter part, <partinfo>K389017</partinfo>
| composite
|
| tightly controlled lac operator, <partinfo>K389050</partinfo>
| composite
|
| Kanamycin-resistance cassette, <partinfo>P1003</partinfo>
| from registry
|
| ccdB-gene, <partinfo>P1010</partinfo>
| from registry
|
| high-copy plasmid with Amp and Tet resistance, <partinfo>pSB1AT3</partinfo>
| from registry
|
| high-copy plasmid with Cm resistance, <partinfo>pSB1C3</partinfo>
| from registry
|
| lac operator, <partinfo>R1010</partinfo>
| from registry
|
|
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