• this contains two <partinfo>B0010</partinfo> terminators
• we used this because of problems mentioned in registry with <partinfo>B0012</partinfo>

• we always assembled a strong RBS (AGGAG)

• this is the lac operon repressor lacI
• used this to build a tightly regulated lac-operon <partinfo>K389050</partinfo> with low basal transcription

• this BioBrick was used as a reporter gene, e.g. in the construct <partinfo>K389013</partinfo>

• this is a strong constitutive promoter which is located upstream of the lacI gene in E. coli lacIq strains
• used this to build the tightly regulated lac-operon <partinfo>K389050</partinfo>

• sensitivity tuner 1...

• sensitivity tuner 2...

• sensitivity tuner 3...

• this BioBrick was used as a visible selection marker for cloning other BioBricks into the <partinfo>pSB1C3</partinfo> plasmid to submit them to the registry

• we always used a medium strong constitutive promoter from the registry to express our BioBricks, so they are neither expressed too strong so the cell suffers from metabolomic stress nor too weak so there is no visible effect of our BioBricks
• this BioBrick was used e.g. in <partinfo>K389010</partinfo> or <partinfo>K389011</partinfo>

• this BioBrick was used to create a plasmid without ColE1 ori
• the R6K ori only works in pir+ / pir116 E. coli strains, so it is used for a two plasmid screening system (other compatibility class than ColE1 ori, easy separation of the two plasmids)
• the R6K ori is a medium copy plasmid in pir+ / pir116 E. coli strains like EC100D

• we wanted to use this 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>

• same as the virA BioBrick - this one from the registry does not work properly, so we made a vir-promoter by ourselves (<partinfo>K389003</partinfo>)
• 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)

• "our" virA
• illegal PstI restriction site removed by site-directed mutagenesis
• isolated from A. tumefaciens C58 TI-plasmid
• functionality of this BioBrick is prooved with <partinfo>K389015</partinfo> and <partinfo>K389016</partinfo>

This is the VirG response regulator from the VirA/G receptor system. The VirA/G receptor system is used by A. tumefaciens to detect phenolic substances secreted by injured plants, e.g. acetosyringone. In presence of these substances, VirG is activated by the VirA receptor and induces the transcription of genes under the control of a vir promoter. Normally, VirG needs the rpoA gene (RNA polymerase subunit) from A. tumefaciens to work in Escherichia coli but this virG BioBrick is mutated, so it works with the rpoA subunit from E. coli. For this reason the point mutations G56V and I77V were brought into the gene (compare YC Jung et al., 2004). Because this BioBrick is synthesized (Mr.Gene GmbH), codon usage is optimized for E. coli and illegal restriction sites were removed. When you use this virG gene in a VirA/G signaling system you do not need <partinfo>BBa_K238010</partinfo> anymore to get the system working in E. coli.

• "our" vir-promoter
• is induced by phosphorylated VirG
• isolated from A. tumefaciens C58 TI-plasmid

• "our" luciferase
• sensitive reporter gene
• isolated from Promega's pGL4.10luc2 plasmid

• "our" kanamycin / neomycin resistance
• used for directed evolution of virA and mutated virA screenings, respectively
• isolated from the BioBrick <partinfo>P1003</partinfo>

• medium strong constitutive promoter, so there is enough VirA receptor expressed but not too much so the cell suffers from the expression
• this version exists in a <partinfo>pSB1C3</partinfo> and a <partinfo>pSB1AT3</partinfo> vector

• the new virG which works without rpoA is expressed constitutively and a kanamycin resistance gene is 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
• this device has to be on a plasmid with different ori than the virA which should be screened (different compatibility classes)
• we cloned this BioBrick into a plasmid with R6K ori which only works in pir+ or pir116 strains (e.g. E. coli EC100D), so we can easily seperate the two plasmids by transforming them into strains without Pir protein (e.g. E. coli TOP10)

• this BioBrick is similar to <partinfo>K389011</partinfo> but with the reporter luciferase instead of an antibiotic resistance gene
• with this BioBrick it is possible to measure the activity of the vir promoter and the VirA receptor, respectively

• this BioBrick is similar to <partinfo>K389011</partinfo> but with the reporter mRFP instead of an antibiotic resistance gene
• with this BioBrick it is possible to measure the activity of the vir promoter and the VirA receptor, respectively

• this BioBrick is an assembly of <partinfo>K389010</partinfo> and <partinfo>K389011</partinfo>
• used for testing the general screening concept of expressing an antibiotic resistance with a vir promoter after induction with acetosyringone (the natural VirA inductor)

• this BioBrick is an assembly of <partinfo>K389010</partinfo> and <partinfo>K389012</partinfo>
• used for characterizing the natural VirA/G system and the vir promoter, respectively

• this BioBrick is an assembly of <partinfo>K389010</partinfo> and <partinfo>K389013</partinfo>
• used for characterizing the natural VirA/G system and the vir promoter, respectively

• this is a complete VirA/G signaling system without a reporter gene, so any gene of interest can be assembled to a vir promoter

• a lac operator is combined with a lacIq so there is always enough lacI to repress the lac operator
• the system is still inducible with IPTG but it has a low basal transcription
• this part is used to test the sensitivity of the luciferase BioBrick <partinfo>K389004</partinfo> and compare it to the sensitivity of the mRFP BioBrick <partinfo>E1010</partinfo>

• a lac operator is combined with a lacIq so there is always enough lacI to repress the lac operator
• the system is still inducible with IPTG but it has a low basal transcription
• ...

• a lac operator is combined with a lacIq so there is always enough lacI to repress the lac operator
• the system is still inducible with IPTG but it has a low basal transcription
• ...

• this is a modified <partinfo>I746370</partinfo> sensitivity tuner
• we erased the arabinose promoter and the GFP readout from the original part and added our firefly luciferase BioBrick as a new readout
• this part can be used behind a random promoter and gives an enhanced luciferase readout signal

• this is a modified <partinfo>I746380</partinfo> sensitivity tuner
• we erased the arabinose promoter and the GFP readout from the original part and added our firefly luciferase BioBrick as a new readout
• this part can be used behind a random promoter and gives an enhanced luciferase readout signal

• this is a modified <partinfo>I746390</partinfo> sensitivity tuner
• we erased the arabinose promoter and the GFP readout from the original part and added our firefly luciferase BioBrick as a new readout
• this part can be used behind a random promoter and gives an enhanced luciferase readout signal

• this is a modified <partinfo>I746370</partinfo> sensitivity tuner + luciferase readout behind a vir promoter
• this part also contains a virG gene under the control of a constitutive promoter
• in front of this part a mutated virA from a <partinfo>K389010</partinfo>-like BioBrick is assembled to get a strong readout signal in order to our goal to make spiciness visible

• this is a modified <partinfo>I746380</partinfo> sensitivity tuner + luciferase readout behind a vir promoter
• this part also contains a virG gene under the control of a constitutive promoter
• in front of this part a mutated virA from a <partinfo>K389010</partinfo>-like BioBrick is assembled to get a strong readout signal in order to our goal to make spiciness visible

• this is a modified <partinfo>I746390</partinfo> sensitivity tuner + luciferase readout behind a vir promoter
• this part also contains a virG gene under the control of a constitutive promoter
• in front of this part a mutated virA from a <partinfo>K389010</partinfo>-like BioBrick is assembled to get a strong readout signal in order to our goal to make spiciness visible

• this is a modified <partinfo>I746370</partinfo> sensitivity tuner + luciferase readout behind a vir promoter characterisation BioBrick
• this part also contains a virG and an unmutated virA gene under the control of a constitutive promoter
• this BioBrick is used to proove the function of the modified sensitivity tuner and to check whether the sensitivity tuner enhances the luciferase signal enough to make it visible

• this is a modified <partinfo>I746380</partinfo> sensitivity tuner + luciferase readout behind a vir promoter characterisation BioBrick
• this part also contains a virG and an unmutated virA gene under the control of a constitutive promoter
• this BioBrick is used to proove the function of the modified sensitivity tuner and to check whether the sensitivity tuner enhances the luciferase signal enough to make it visible

• this is a modified <partinfo>I746390</partinfo> sensitivity tuner + luciferase readout behind a vir promoter characterisation BioBrick
• this part also contains a virG and an unmutated virA gene under the control of a constitutive promoter
• this BioBrick is used to proove the function of the modified sensitivity tuner and to check whether the sensitivity tuner enhances the luciferase signal enough to make it visible

• we used this BioBrick to create <partinfo>K389005</partinfo>

• we used this BioBrick for 3A-assemblies

• we used this plasmids in 3A-assemblies and as the backbone of the mutated and unmutated <partinfo>K389010</partinfo> part

• the plasmid to send in the BioBricks

• used to build a tightly regulated lac operator (<partinfo>K389050</partinfo>)