Team:Bielefeld-Germany/Results/Used

From 2010.igem.org

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|style="border-style: solid; border-width: 1px"| ''vir'' promoter part, <partinfo>K389017</partinfo>
|style="border-style: solid; border-width: 1px"| ''vir'' promoter part, <partinfo>K389017</partinfo>
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|style="border-style: solid; border-width: 1px"| tightly controlled ''lac'' operator, <partinfo>K389050</partinfo>
|style="border-style: solid; border-width: 1px"| tightly controlled ''lac'' operator, <partinfo>K389050</partinfo>
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|style="border-style: solid; border-width: 1px"| Kanamycin-resistance cassette, <partinfo>P1003</partinfo>
|style="border-style: solid; border-width: 1px"| Kanamycin-resistance cassette, <partinfo>P1003</partinfo>
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|style="border-style: solid; border-width: 1px"| ''ccdB''-gene, <partinfo>P1010</partinfo>
|style="border-style: solid; border-width: 1px"| ''ccdB''-gene, <partinfo>P1010</partinfo>
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|style="border-style: solid; border-width: 1px"| high-copy plasmid with Amp and Tet resistance, <partinfo>pSB1AT3</partinfo>
|style="border-style: solid; border-width: 1px"| high-copy plasmid with Amp and Tet resistance, <partinfo>pSB1AT3</partinfo>
|style="border-style: solid; border-width: 1px"| from registry
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|style="border-style: solid; border-width: 1px"| high-copy plasmid with Cm resistance, <partinfo>pSB1C3</partinfo>
|style="border-style: solid; border-width: 1px"| high-copy plasmid with Cm resistance, <partinfo>pSB1C3</partinfo>
|style="border-style: solid; border-width: 1px"| from registry
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|style="border-style: solid; border-width: 1px"| ''lac'' operator, <partinfo>R0010</partinfo>
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Revision as of 14:03, 28 September 2010

{{{1}}}

Used BioBricks

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>R0010</partinfo> from registry
  • ...
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