Team:LMU-Munich/Data/Vectors
From 2012.igem.org
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For the detailed protocols, please see our [https://2012.igem.org/Team:LMU-Munich/Lab_Notebook/Protocols protocol section]. | For the detailed protocols, please see our [https://2012.igem.org/Team:LMU-Munich/Lab_Notebook/Protocols protocol section]. | ||
+ | <div class="box"> | ||
+ | ===Overview=== | ||
+ | {| class="colored" | ||
+ | !Name BioBrick | ||
+ | !''E. coli'' res. | ||
+ | !''B. subt.'' res. | ||
+ | !Insertion locus | ||
+ | !Description | ||
+ | !Cloning status | ||
+ | !Works? | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>1C [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823023 <font color="#EBFCE4" size="2">(BBa_K823023)</font>] | ||
+ | |Amp | ||
+ | |Cm | ||
+ | |amyE | ||
+ | |empty | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>4S [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823022 <font color="#EBFCE4" size="2">(BBa_K823022)</font>] | ||
+ | |Amp | ||
+ | |Spec | ||
+ | |thrC | ||
+ | |empty | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>2E [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823027 <font color="#EBFCE4" size="2">(BBa_K823027)</font>] | ||
+ | |Amp | ||
+ | |MLS | ||
+ | |lacA | ||
+ | |empty | ||
+ | |in work | ||
+ | |not tested | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>1C-<i>lac</i>Z [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823021 <font color="#EBFCE4" size="2">(BBa_K823021) </font>] | ||
+ | |Amp | ||
+ | |Cm | ||
+ | |amyE | ||
+ | |with <i>lacZ</i> reporter gene | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>3C-<i>lux</i>ABCDE [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823025 <font color="#EBFCE4" size="2">(BBa_K823025)</font>] | ||
+ | |Amp | ||
+ | |Cm | ||
+ | |sacA | ||
+ | |with <i>luxABCDE</i> reporter cassette | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>4S-P<sub><i>Xyl</i></sub> [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823024 <font color="#EBFCE4" size="2">(BBa_K823024)</font>] | ||
+ | |Amp | ||
+ | |Spec | ||
+ | |thrC | ||
+ | |with Xylose-inducible promoter | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |unexpected | ||
+ | |- | ||
+ | !pSB<sub>Bs</sub>0K-P<sub><i>spac</i></sub> [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823026 <font color="#EBFCE4" size="2">(BBa_K823026)</font>] | ||
+ | |Amp | ||
+ | |Kan | ||
+ | |replicative | ||
+ | |with IPTG inducible promoter | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |refuses transformation | ||
+ | |- | ||
+ | |} | ||
+ | |[[File:BoxUnchecked.png|40px|left]] | ||
+ | |[[File:BoxChecked.png|40px|left]] | ||
+ | |||
+ | </div> | ||
<div class="box"> | <div class="box"> | ||
===pSB<sub>Bs</sub>1C=== | ===pSB<sub>Bs</sub>1C=== |
Revision as of 15:24, 21 September 2012
![Team-LMU culture tubes.resized.jpg](/wiki/images/d/d8/Team-LMU_culture_tubes.resized.jpg)
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
[ more news ]
![Sporenfreunde](https://static.igem.org/mediawiki/2012/e/e3/Team_LMU_Sporenfreunde.png)
Bacillus subtilis vectors
For the detailed protocols, please see our protocol section.
Overview
Name BioBrick | E. coli res. | B. subt. res. | Insertion locus | Description | Cloning status | Works? |
---|---|---|---|---|---|---|
pSBBs1C [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823023 (BBa_K823023)] | Amp | Cm | amyE | empty | ||
pSBBs4S [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823022 (BBa_K823022)] | Amp | Spec | thrC | empty | ||
pSBBs2E [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823027 (BBa_K823027)] | Amp | MLS | lacA | empty | in work | not tested |
pSBBs1C-lacZ [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823021 (BBa_K823021) ] | Amp | Cm | amyE | with lacZ reporter gene | ||
pSBBs3C-luxABCDE [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823025 (BBa_K823025)] | Amp | Cm | sacA | with luxABCDE reporter cassette | ||
pSBBs4S-PXyl [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823024 (BBa_K823024)] | Amp | Spec | thrC | with Xylose-inducible promoter | unexpected | |
pSBBs0K-Pspac [http://partsregistry.org/wiki/index.php?title=Part:BBa_K823026 (BBa_K823026)] | Amp | Kan | replicative | with IPTG inducible promoter | refuses transformation |
pSBBs1C
pSBBs1C is one of our empty vectors. It integrates into the amyE locus of Bacillus subtilis and carries a Chloramphenicol resistance as well as an Ampicillin resistance for E. coli. Inbetween the two recombination sites, there is the constitutively expressed Cm resistance and the multiple cloning site which contains RFP to simplify the insertion of your BioBrick.The plasmid is then transformed into B. subtilis and checked via the resistance for "integration at all" and via a starch test for "integration into the right locus". Please note that the colonies without a bright surounding are correct.
We tested this vector with various inserts, for example the final construct of our Sporobeads with GFP. The starch test shows that the insertion in in the amyE locus and fluorescence microscopy reveals the functionality of the construct.
pSBBs4S
pSBBs4S also is an empty vector with only the multiple cloning site and the spectinomycine resistence gene in between the homologous regions. This vector integrates into the thrC locus of B. subtilis.
We tested the integration of this plasmid with the threonine test and a construct from the Suicideswitch as insert.