IGEM:IMPERIAL/2008/Prototype/Wetlab/parts/Composite

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Composite Parts

Seen as how the wiki may soon cause errors due to the size of the previous page (and the affect it was having on us), here is the new parts page, specifically for composite parts...

We'll start looking at these soon and I'll get back to people after the weekend

Expression Related

P43-gsiB (done)

Name: Promoter 43 and RBS gsiB for B. subtilis

Code: K143050

Long: Constitutive promoter 43(<bbpart>BBa_K143013</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

P43-gsiB can be used in the context of a Ribosomes per second (PoPS) output generator

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of P43 and RBS-gsiB were obtained from papers[1, 2] and the sequence synthesised by GeneArt

Source: P43-gsiB was synthesised by GeneArt

References

Error fetching PMID 16000826:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 16000826: [1]
  3. Error fetching PMID 9669336: [2]
All Medline abstracts: PubMed HubMed

P43-spoVG (done)

Name: Promoter 43 and RBS spoVG for B. subtilis

Code: K143051

Long: Constitutive promoter 43(<bbpart>BBa_K143013</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

P43-spoVG can be used in the context of a Ribosomes per seconds (PoPS) output generator

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of P43 and RBS-spoVG were obtained from papers[1, 2] and the sequence synthesised by GeneArt

Source: P43-spoVG was synthesised by GeneArt

References

Error fetching PMID 16000826:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 16000826: [1]
  3. Error fetching PMID 9669336: [2]
All Medline abstracts: PubMed HubMed

Pveg-gsiB (done)

Name: Promoter Pveg and RBS gsiB for B. subtilis

Code: K143052

Long: Constitutive promoter veg(<bbpart>BBa_K143012</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

Pveg-gsiB can be used in the context of a Ribosomes per second (RiPS) output generator

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of Pveg was obtained from the DBTBS[1] and RBS-gsiB were obtained from papers[2] and the sequence synthesised by GeneArt

Source: Pveg-gsiB was synthesised by GeneArt

References

Error fetching PMID 17962296:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 17962296: [1]
  3. Error fetching PMID 9669336: [2]
All Medline abstracts: PubMed HubMed

Pveg-spoVG (Done)

Name: Promoter Pveg and RBS spoVG for B. subtilis

Code: K143053

Long: Constitutive promoter veg(<bbpart>BBa_K143012</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

Pveg-spoVG can be used in the context of a Ribosomes per second (RiPS) output generator

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of Pveg was obtained from the DBTBS[1] and RBS-spoVG were obtained from papers[2] and the sequence synthesised by GeneArt

Source: Pveg-spoVG was synthesised by GeneArt

References

Error fetching PMID 17962296:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 17962296: [1]
  3. Error fetching PMID 9669336: [2]
All Medline abstracts: PubMed HubMed


Phyper-spank-gsiB (Done)

Name: Promoter hyper-spank and RBS gsiB for B. subtilis

Code: K143054

Long: Inducible promoter hyper-spank(<bbpart>BBa_K143015</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

Phyperspank-gsiB can be used to take an input of IPTG and give a Ribosomes per second (RiPS) output generator.

IPTG does not directly induce the expression of the promoter hyper-spank, but requires the transcriptional regulator LacI, (<bbpart>BBa_K143035</bbpart>). This means that LacI must be constitutively expressed in B.subtilis in order to use the promoter hyper-spank as an inducible promoter.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter hyperspank was obtained from the B. subtilis integration vector pDR111 and RBS-gsiB were obtained from papers[1] and the sequence synthesised by GeneArt

Source: Phyperspank-gsiB was synthesised by GeneArt

References

Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 9669336: [1]


Phyper-spank-spoVG (Done)

Name: Promoter hyper-spank and RBS spoVG for B. subtilis

Code: K143055

Long: Inducible promoter hyper-spank(<bbpart>BBa_K143015</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

Phyperspank-spoVG can be used to take an input of IPTG and give a Ribosomes per second (RiPS) output generator.

IPTG does not directly induce the expression of the promoter hyper-spank, but requires the transcriptional regulator LacI, (<bbpart>BBa_K143033</bbpart>). This means that LacI must be constitutively expressed in B.subtilis in order to use the promoter hyper-spank as an inducible promoter.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter hyperspank was obtained from the B. subtilis integration vector pDR111 and RBS-spoVG were obtained from papers[1] and the sequence synthesised by GeneArt

Source: Phyperspank-spoVG was synthesised by GeneArt

References

Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 9669336: [1]

Pxyl-gsiB (Done)

Name: Promoter xyl and RBS gsiB for B. subtilis

Code: K143056

Long: Inducible promoter xyl(<bbpart>BBa_K143014</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

Pxyl-gsiB can be used to take an input of xylose and give a Ribosomes per second (RiPS) output generator.

Xylose does not directly induce the expression of the promoter xyl, but requires the transcriptional regulator XylR, (<bbpart>BBa_K143036</bbpart>). This means that XylR must be constitutively expressed in B.subtilis in order to use the promoter hyper-spank as an inducible promoter. XylR is naturally expressed by B. subtilis but should be upregulated to increase efficiency.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter xyl and RBS-gsiB were obtained from papers[1, 2, 3] and the sequence synthesised by GeneArt

Source: Pxyl-gsiB was synthesised by GeneArt

References

Error fetching PMID 8973310:
Error fetching PMID 11274134:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 8973310: [1]
  3. Error fetching PMID 11274134: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed

Pxyl-spoVG (Done)

Name: Promoter xyl and RBS spoVG for B subtilis

Code: K143057

Long:Inducible promoter xyl(<bbpart>BBa_K143014</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

Pxyl-spoVG can be used to take an input of xylose and give a Ribosomes per second (RiPS) output generator.

Xylose does not directly induce the expression of the promoter xyl, but requires the transcriptional regulator XylR, (<bbpart>BBa_K143036</bbpart>). This means that XylR must be constitutively expressed in B.subtilis in order to use the promoter hyper-spank as an inducible promoter. XylR is naturally expressed by B. subtilis but should be upregulated to increase efficiency.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter xyl and RBS-spoVG were obtained from papers[1, 2, 3] and the sequence synthesised by GeneArt

Source: Pxyl-spoVG was synthesised by GeneArt

References

Error fetching PMID 8973310:
Error fetching PMID 11274134:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 8973310: [1]
  3. Error fetching PMID 11274134: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed

Pctc-gsiB (Done)

Name Promoter ctc and RBS gsiB for B. subtilis

Code: K143058

Long: Sigma B dependant promoter ctc(<bbpart>BBa_K143010</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

In B. subtilis endogenous sigma factor B is activated under mild stress. These mild stress conditions can be generally split into nutrient stress response and physical stress response. Nutrient stress response is triggered by low levels of ATP and GTP and physical stress response is triggered by exposure to blue light, salt, heat, acid or ethanol[1]. The promoter ctc has been used previously as a read out for the activation of sigma factor B [2].

Pctc has been used to take an input of blue light and give a Ribosomes per second(RiPS) output. To work as a blue light receiver correctly, over-expression of the blue light receptor YtvA (<bbpart>BBa_K143037</bbpart>) is required.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter ctc was obtained from the B. subtilis genome and published papers[1, 2]. RBS-gsiB were obtained from papers[3] and the sequence synthesised by GeneArt

Source: Pctc-gsiB was synthesised by GeneArt

References

Error fetching PMID 3100810:
Error fetching PMID 17575448:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 3100810: [1]
  3. Error fetching PMID 17575448: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed


Pctc-spoVG (Done)

Name: Promoter ctc and RBS spoVG for B. subtilis

Code: K143059

Long: Sigma B dependant promoter ctc(<bbpart>BBa_K143010</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

In B. subtilis endogenous sigma factor B is activated under mild stress. These mild stress conditions can be generally split into nutrient stress response and physical stress response. Nutrient stress response is triggered by low levels of ATP and GTP and physical stress response is triggered by exposure to blue light, salt, heat, acid or ethanol[1]. The promoter ctc has been used previously as a read out for the activation of sigma factor B [2].

Pctc has been used to take an input of blue light and give a Ribosomes per second(RiPS) output. To work as a blue light receiver correctly, over-expression of the blue light receptor YtvA (<bbpart>BBa_K143037</bbpart>) is required.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter ctc was obtained from the B. subtilis genome and published papers[1, 2]. RBS-spoVG were obtained from papers[3] and the sequence synthesised by GeneArt

Source: Pctc-spoVG was synthesised by GeneArt

References

Error fetching PMID 3100810:
Error fetching PMID 17575448:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 3100810: [1]
  3. Error fetching PMID 17575448: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed


PgsiB-gsiB (Done)

Name: Promoter gsiB and RBS gsiB for B. subtilis

Code: K143060

Long: Sigma B dependant promoter gsiB(<bbpart>BBa_K143011</bbpart>) coupled to the strong Ribosome Binding Site gsiB(<bbpart>BBa_K143020</bbpart>) from B. subtilis.

In B. subtilis endogenous sigma factor B is activated under mild stress. These mild stress conditions can be generally split into nutrient stress response and physical stress response. Nutrient stress response is triggered by low levels of ATP and GTP and physical stress response is triggered by exposure to blue light, salt, heat, acid or ethanol[1]. The promoter ctc has been used previously as a read out for the activation of sigma factor B [2].

PgsiB has been used to take an input of blue light and give a Ribosomes per second(RiPS) output. To work as a blue light receiver correctly, over-expression of the blue light receptor YtvA (<bbpart>BBa_K143037</bbpart>) is required.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter gsiB was obtained from the B. subtilis genome and published papers[1, 2]. RBS-gsiB were obtained from papers[3] and the sequence synthesised by GeneArt

Source: PgsiB-gsiB was synthesised by GeneArt

References

Error fetching PMID 3100810:
Error fetching PMID 17575448:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 3100810: [1]
  3. Error fetching PMID 17575448: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed

PgsiB-spoVG (Done)

Name: Promoter gsiB and RBS spoVG for B. subtilis

Code: K143061

Long: Sigma B dependant promoter gsiB(<bbpart>BBa_K143011</bbpart>) coupled to the strong Ribosome Binding Site spoVG(<bbpart>BBa_K143021</bbpart>) from B. subtilis.

In B. subtilis endogenous sigma factor B is activated under mild stress. These mild stress conditions can be generally split into nutrient stress response and physical stress response. Nutrient stress response is triggered by low levels of ATP and GTP and physical stress response is triggered by exposure to blue light, salt, heat, acid or ethanol[1]. The promoter ctc has been used previously as a read out for the activation of sigma factor B [2].

PgsiB has been used to take an input of blue light and give a Ribosomes per second(RiPS) output. To work as a blue light receiver correctly, over-expression of the blue light receptor YtvA (<bbpart>BBa_K143037</bbpart>) is required.

To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix[1]. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency.

Design: The sequence of promoter gsiB was obtained from the B. subtilis genome and published papers[1, 2]. RBS-spoVG were obtained from papers[3] and the sequence synthesised by GeneArt

Source: Pctc-spoVG was synthesised by GeneArt

References

Error fetching PMID 3100810:
Error fetching PMID 17575448:
Error fetching PMID 9669336:
  1. http://partsregistry.org/Assembly:RBS-CDS_issues

    [1]

  2. Error fetching PMID 3100810: [1]
  3. Error fetching PMID 17575448: [2]
  4. Error fetching PMID 9669336: [3]
All Medline abstracts: PubMed HubMed

LacI-Terminator (Done)

Name: LacI repressor protein - Terminator

Code: K143062

Long: LacI transcriptional repressor protein (<bbpart>BBa_K143033</bbpart>) coupled to the double terminator (<bbpart>BBa_B0015</bbpart>.

The LacI does not possess a LVA degradation tag and gas a short (3 amino acid) N-terminal deletion consistent with LacI used in conjunction with B. subtilis.

LacI can be used in conjunction with the lac operon promoter (<bbpart>BBa_K143015</bbpart>), where the LacI will act as a receiver for an IPTG input to result in a Polymerases per second (PoPS) output.

The double terminator is the most commonly used terminator and is a combination of parts <bbpart>BBa_B0010</bbpart> and <bbpart>BBa_B0012</bbpart>.

The double terminator allows the LacI to be easily incorporated into a closed transcriptional unit.

Design: LacI was identified from the pDR111 B. subtilis integration vector. The double terminator is the most commonly used registry terminator.

Source: LacI was produced by PCR cloning using Pfu form the B. subtilis integration vector and cloned into a BioBrick with the registry double terminator

XylR-Terminator (Done)

Name: Xylose operon repressor protein - Terminator

Code: K143063

Long: Xylose operon repressor protein(XylR)(<bbpart>BBa_K143036</bbpart>) coupled to the double terminator (<bbpart>BBa_B0015</bbpart>.

XylR can be used in conjunction with the xylose operon promoter (<bbpart>BBa_K143014</bbpart>), where the XylR will act as a receiver for a xylose input to result in a Polymerases per second (PoPS) output.

The double terminator is the most commonly used terminator and is a combination of parts <bbpart>BBa_B0010</bbpart> and <bbpart>BBa_B0012</bbpart>.

The double terminator allows the XylR to be easily incorporated into a closed transcriptional unit.

Design: XylR was identified from the B. subtilis chromosome. The double terminator is the most commonly used registry terminator.

Source: XylR was produced by PCR cloning using Pfu form the B. subtilis chromosome and cloned into a BioBrick with the registry double terminator

YtvA-Terminator (Done)

Name: YtvA light receptor Protein - Terminator

Code: K143066

Long: YtvA light receptor protein (YtvA)(<bbpart>BBa_K143037</bbpart>) coupled to the double terminator (<bbpart>BBa_B0015</bbpart>).

YtvA can be used in conjunction with the sigma factor B specific promoter (<bbpart>BBa_K143010</bbpart> or <bbpart>BBa_K143011</bbpart>), where the YtvA will act as a receiver for a blue light input to result in a Polymerases per second (PoPS) output.

The double terminator is the most commonly used terminator and is a combination of parts <bbpart>BBa_B0010</bbpart> and <bbpart>BBa_B0012</bbpart>.

The double terminator allows the YtvA to be easily incorporated into a closed transcriptional unit.

Design: YtvA was identified from the B. subtilis chromosome. The double terminator is the most commonly used registry terminator.

Source: YtvA was synthesised by GeneArt and cloned into a BioBrick with the registry double terminator

Antibiotic Related

Chloraphemicol Resistance Protein - Terminator (Done)

Name: Chloraphemicol resistance protein - Terminator

Code: K143064

Long: Chloraphemicol acetyltransferase protein(<bbpart>BBa_J31005</bbpart>) coupled to the double terminator (<bbpart>BBa_B0015</bbpart>).

Chloraphemicol acetyltransferase confers resistance to Chloraphemicol

The double terminator is the most commonly used terminator and is a combination of parts <bbpart>BBa_B0010</bbpart> and <bbpart>BBa_B0012</bbpart>.

The double terminator allows the CAT to be incorporated into a closed transcriptional unit.

Design: Chloraphemicol acetyltransferase is an exisiting registry protein. The double terminator is the most commonly used registry termiantor

Source: The Chloraphemicol acetyltransferase and double terminator were taken both taken from the registry.

Spectinomycin Resistance Protein (Aad9) - Terminator (Done)

Name: Spectinomycin Resistance Protein (Aad9) - Terminator

Code: K143065

Long: Aad9 spectinomycin resistance protein(<bbpart>BBa_</bbpart>) coupled to the double terminator (<bbpart>BBa_B0015</bbpart>).

Aad9 confers resistance to spectinomycin.

The double terminator is the most commonly used terminator and is a combination of parts <bbpart>BBa_B0010</bbpart> and <bbpart>BBa_B0012</bbpart>.

The double terminator allows the Spectinomycin resistance gene to be incorporated into a closed transcriptional unit.

Design: Aad9 was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase. The double terminator is the most commonly used registry termiantor.

Source: Aad9 was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick iwth the double terminator was taken from the registry

Integration Related

AmyE integratable PoPS generator (P43-gsiB) (Partially Uploaded)

Name: AmyE integratable PoPS generator (P43-gsiB)

Code: K143067

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to the PoPS generator P43-gsiB (<bbpart>BBa_K143050</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The P43-gsiB promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of P43-gsiB was obtained from papers.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the P43-gsiB that was synthesised by GeneArt

AmyE integratable PoPS generator (P43-spoVG) (Partially Uploaded)

Name: AmyE integratable PoPS generator (P43-spoVG)

Code: K143068

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to the PoPS generator P43-spoVG (<bbpart>BBa_K143051</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The P43-spoVG promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of P43-spoVG was obtained from papers.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the P43-spoVG that was synthesised by GeneArt

AmyE integratable PoPS generator (Pveg-gsiB) (Partially Uploaded)

Name: AmyE integratable PoPS generator (Pveg-gsiB)

Code: K143069

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to the PoPS generator Pveg-gsiB (<bbpart>BBa_K143052</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The Pveg-gsiB promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of Pveg-gsiB was obtained from papers.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the Pveg-gsiB that was synthesised by GeneArt

AmyE integratable PoPS generator (Pveg-spoVG) (Ready for Upload)

Name: AmyE integratable PoPS generator (Pveg-spoVG)

Code: K143070

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to the PoPS generator Pveg-spoVG (<bbpart>BBa_K143053</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The Pveg-spoVG promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of Pveg-spoVG was obtained from papers.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the Pveg-spoVG that was synthesised by GeneArt

AmyE integratable PoPS generator (P43-gsiB) (with CmR) (Ready for Upload)

Name: AmyE integratable PoPS generator (P43-gsiB) (with CmR)

Code: K143071

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to a chloramphenicol resistance generator in closed transcriptional unit (Parts <bbpart>BBa_K143050</bbpart> and <bbpart>BBa_K143064</bbpart>) and the PoPS generator P43-gsiB (<bbpart>BBa_K143050</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The chloramphenicol adenyltransferase give resistance to chloramphenicol while the terminator prevents readthrough.

The P43-gsiB promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of P43-gsiB was obtained from papers. Chloramphenicol adenyltransferase and the double terminator were obtained from the registry.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the P43-gsiB that was synthesised by GeneArt and Chloramphenicol adenyltransferase and the double terminator that were obtained from the registry.

AmyE integratable PoPS generator (P43-spoVG) (with CmR) (Ready for Upload)

Name: AmyE integratable PoPS generator (P43-spoVG) (with CmR)

Code: K143072

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to a chloramphenicol resistance generator in closed transcriptional unit (Parts <bbpart>BBa_K143051</bbpart> and <bbpart>BBa_K143064</bbpart>) and the PoPS generator P43-spoVG (<bbpart>BBa_K143051</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The chloramphenicol adenyltransferase give resistance to chloramphenicol while the terminator prevents readthrough.

The P43-spoVG promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of P43-spoVG was obtained from papers. Chloramphenicol adenyltransferase and the double terminator were obtained from the registry.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the P43-spoVG that was synthesised by GeneArt and Chloramphenicol adenyltransferase and the double terminator that were obtained from the registry.

AmyE integratable PoPS generator (Pveg-gsiB) (with CmR) (Ready for Upload)

Name: AmyE integratable PoPS generator (Pveg-gsiB) (with CmR)

Code: K143073

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to a chloramphenicol resistance generator in closed transcriptional unit (Parts <bbpart>BBa_K143052</bbpart> and <bbpart>BBa_K143064</bbpart>) and the PoPS generator Pveg-gsiB (<bbpart>BBa_K143052</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The chloramphenicol adenyltransferase give resistance to chloramphenicol while the terminator prevents readthrough.

The Pveg-gsiB promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of Pveg-gsiB was obtained from papers. Chloramphenicol adenyltransferase and the double terminator were obtained from the registry.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the Pveg-gsiB that was synthesised by GeneArt and Chloramphenicol adenyltransferase and the double terminator that were obtained from the registry.

AmyE integratable PoPS generator (Pveg-spoVG) (with CmR) (Ready for Upload)

Name: AmyE integratable PoPS generator (Pveg-spoVG) (with CmR)

Code: K143074

Long: AmyE 5' Integration sequence(<bbpart>BBa_K143001</bbpart>) coupled to a chloramphenicol resistance generator in closed transcriptional unit (Parts <bbpart>BBa_K143053</bbpart> and <bbpart>BBa_K143064</bbpart>) and the PoPS generator Pveg-spoVG (<bbpart>BBa_K143053</bbpart>).

The amyE 5' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 3' amyE integration sequence(<bbpart>BBa_K143002</bbpart>) is cloned onto the 3' end of the construct.

The chloramphenicol adenyltransferase give resistance to chloramphenicol while the terminator prevents readthrough.

The Pveg-spoVG promoter and RBS for B. subtilis constitutively generate a PoPS output.

Design: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. The sequence of Pveg-spoVG was obtained from papers. Chloramphenicol adenyltransferase and the double terminator were obtained from the registry.

Source: The amyE 5' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the Pveg-spoVG that was synthesised by GeneArt and Chloramphenicol adenyltransferase and the double terminator that were obtained from the registry.

B. subtilis AmyE locus GFP promoter and RBS tester (Ready for Upload)

Name: B. subtilis AmyE locus GFP output promoter and RBS tester

Code: K143075

Long: AmyE 3' Integration sequence(<bbpart>BBa_K143002</bbpart>) coupled to GFP proein coding region and the registry double terminator (part <bbpart>BBa_I13401</bbpart>.

The amyE 3' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 5' amyE integration sequence(<bbpart>BBa_K143001</bbpart>) is cloned onto the 5' end of the construct.

The GFP coding region gives a fluorescent output when a promoter and RBS are placed immediately upstream of the coding region while the terminator prevents readthrough.

Design: The amyE 3' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. GFP and the double terminator were obtained from the registry.

Source: The amyE 3' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the GFP and the double terminator that were obtained from the registry.

B. subtilis AmyE locus RFP promoter and RBS tester (Ready for Upload)

Name: B. subtilis AmyE locus RFP output promoter and RBS tester

Code: K143076

Long: Long: AmyE 3' Integration sequence(<bbpart>BBa_K143002</bbpart>) coupled to RFP proein coding region and the registry double terminator (part <bbpart>BBa_J04650</bbpart>.

The amyE 3' integration sequence allows integration into the B. subtilis genome at the amyE locus if the 5' amyE integration sequence(<bbpart>BBa_K143001</bbpart>) is cloned onto the 5' end of the construct.

The RFP coding region gives a fluorescent output when a promoter and RBS are placed immediately upstream of the coding region while the terminator prevents readthrough.

Design: The amyE 3' integration sequence was PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase. RFP and the double terminator were obtained from the registry.

Source: The amyE 3' integration sequence was PCR cloned from the B. subtilis integration vector utilising Pfu DNA polymerase and cloned into a BioBrick with the RFP and the double terminator that were obtained from the registry.

P43-gsiB GFP expression construct (Ready for Upload)

Name: P43-gsiB GFP expression construct

Code: K143077

Long: GFP gene under expresson of the P43 promoter and gsiB RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the P43 promoter and gsiB RBS (Combined part <bbpart>BBa_K143050</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, GFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter P43 and RBS gsiB were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the P43-gsiB part that was synthesised by GeneArt.

P43-spoVG GFP expression construct (Ready for Upload)

Name: P43-spoVG GFP expression construct

Code: K143078

Long: GFP gene under expresson of the P43 promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the P43 promoter and spoVG RBS (Combined part <bbpart>BBa_K143051</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, GFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter P43 and RBS spoVG were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the P43-spoVG part that was synthesised by GeneArt.

Pveg-spoVG GFP expression construct (Ready for Upload)

Name: Pveg-spoVG GFP expression construct

Code: K143079

Long: GFP gene under expresson of the Pveg promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the Pveg promoter and spoVG RBS (Combined part <bbpart>BBa_K143053</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, GFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter Pveg and RBS spoVG were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the Pveg-spoVG part that was synthesised by GeneArt.

P43-gsiB RFP expression construct (Ready for Upload)

Name: P43-gsiB RFP expression construct

Code: K143080

Long: RFP gene under expresson of the P43 promoter and gsiB RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the P43 promoter and gsiB RBS (Combined part <bbpart>BBa_K143050</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, RFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter P43 and RBS gsiB were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the P43-gsiB part that was synthesised by GeneArt.

P43-spoVG RFP expression construct (Ready for Upload)

Name: P43-spoVG RFP expression construct

Code: K143081

Long: RFP gene under expresson of the P43 promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the P43 promoter and spoVG RBS (Combined part <bbpart>BBa_K143051</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, RFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter P43 and RBS spoVG were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the P43-spoVG part that was synthesised by GeneArt.

Pveg-spoVG RFP expression construct (Ready for Upload)

Name: Pveg-spoVG RFP expression construct

Code: K143082

Long: RFP gene under expresson of the Pveg promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.

Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.

This device was used by the Imperial iGEM 2008 team to characterise the Pveg promoter and spoVG RBS (Combined part <bbpart>BBa_K143053</bbpart>) as part of the Biofabricator subtilis project.

Design: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 using Pfu DNA polymerase. The double terminator, RFP gene and chlorapmphenicol acetyltransferase gene were taken from the registry. The sequence of promoter Pveg and RBS spoVG were taken from papers.

Source: The 5' and 3' amyE integration sequences were PCR cloned from the B. subtilis integration vector pDR111 utilising Pfu DNA polymerase and cloned into a BioBrick with the parts from the registry and the Pveg-spoVG part that was synthesised by GeneArt.

Codes and associated parts

Note
  • Aad9 is the Spectinomycin resistance gene
  • RI - Resistance Integration Brick, P - Promoter, Pi - chemically inducible promoter, Pl - light inducible promoter, Bs - B.subtilis, PTC - Promoter Testing Construct, Rep - Repressor protein
Code Part Code Part Code Part Code Part Code Part
K143050 P43-gsiB K143051 P43-spoVG K143052 Pveg-gsiB K143053 Pveg-spoVG K143054 Phyperspank-gsiB
K143055 Phyper-spank-spoVG K143056 Pxyl-gsiB K143057 Pxyl-spoVG K143058 Pctc-gsiB K143059 Pctc-spoVG
K143060 PgsiB-gsiB K143061 PgsiB-spoVG K143062 LacI - Terminator K143063 XylR - Terminator K143064 CAT - Terminator
K143065 Aad9 - Terminator K143066 Ytva - Terminator K143067 5' 9 K143068 5' 10 K143069 5' 11
K143070 5' 12 K143071 RI Brick 9 tail K143072 RI Brick 10 tail K143073 RI Brick 11 tail K143074 RI Brick 12 tail
K143075 GFP - Terminator 3' K143076 RFP - Terminator 3' K143077 P43-gsiB GFP expression construct K143078 P43-spoVG GFP expression construct K143079 Pveg-spoVG GFP expression construct
K143080 P43-gsiB RFP expression construct K143081 P43-spoVG RFP expression construct K143082 Pveg-spoVG RFP expression construct K143083 K143084
K143085 K143086 K143087 K143088 K143089
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