"
Team:Groningen/Parts/Used Parts
From 2009.igem.org
m (→Vectors) |
m (→Used Parts) |
||
| Line 17: | Line 17: | ||
The ligation of a part behind the RBS succeded, and was confirmed by gel (correct vector size) and sequencing with VF2. We used this part in combination with several genes for building our biobricks e.g. <partinfo>BBa_K190061</partinfo>. | The ligation of a part behind the RBS succeded, and was confirmed by gel (correct vector size) and sequencing with VF2. We used this part in combination with several genes for building our biobricks e.g. <partinfo>BBa_K190061</partinfo>. | ||
|} | |} | ||
| - | |||
'''<partinfo>BBa_B0014</partinfo> Double terminator (B0012-B0011)''' <br> Double terminator consisting of BBa_B0012 and BBa_B0011 <br> | '''<partinfo>BBa_B0014</partinfo> Double terminator (B0012-B0011)''' <br> Double terminator consisting of BBa_B0012 and BBa_B0011 <br> | ||
| - | |||
| - | |||
'''<partinfo>BBa_I750016</partinfo> GVP Gas Vesicle Proteins''' <br> This parts creates gas vesicles inside the cell wich enables it to float. It consist of 6064 base pairs. It's backbone is <partinfo>BBa_J61035</partinfo> <br> | '''<partinfo>BBa_I750016</partinfo> GVP Gas Vesicle Proteins''' <br> This parts creates gas vesicles inside the cell wich enables it to float. It consist of 6064 base pairs. It's backbone is <partinfo>BBa_J61035</partinfo> <br> | ||
| - | + | '''<partinfo>BBa_P1010</partinfo> ccdB cell death gene''' | |
| - | + | ||
| - | '''<partinfo>BBa_P1010</partinfo> ccdB cell death gene''' | + | |
{| | {| | ||
| Line 34: | Line 29: | ||
<I> iGEM Groningen 2009 </I> | <I> iGEM Groningen 2009 </I> | ||
|width='60%' valign='top'| | |width='60%' valign='top'| | ||
| - | The ccdB cell death gene worked as expected killing our ''E. coli'' TOP10 cells, and keeping our ''E. coli'' DB3 cells alive. After noticing the inconsistent sequencing result for the pSB2K3 plasmid with ccdB cell death gene, we decided to choose a different pSB2K3 plasmid with random part to continue with our assemblies. This to minimize the chance of unwanted surprises in the end. | + | P1010 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or religated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result. The ccdB cell death gene worked as expected killing our ''E. coli'' TOP10 cells, and keeping our ''E. coli'' DB3 cells alive. After noticing the inconsistent sequencing result for the pSB2K3 plasmid with ccdB cell death gene, we decided to choose a different pSB2K3 plasmid with random part to continue with our assemblies. This to minimize the chance of unwanted surprises in the end. |
|} | |} | ||
| - | |||
| - | |||
===Promotors=== | ===Promotors=== | ||
| - | '''<partinfo>J23100</partinfo> Constitutive promoter family member (high expression)''' <br> pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> | + | '''<partinfo>J23100</partinfo> Constitutive promoter family member (high expression)''' <br> pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> |
| - | '''<partinfo>J23101</partinfo> Constitutive promoter family member (high expression, reference)''' <br> pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> | + | '''<partinfo>J23101</partinfo> Constitutive promoter family member (high expression, reference)''' <br> pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> |
| - | '''<partinfo>J23106</partinfo> Constitutive promoter family member (medium expression)''' <br> pMed is a medium consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> | + | '''<partinfo>J23106</partinfo> Constitutive promoter family member (medium expression)''' <br> pMed is a medium consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo> <br> |
| - | '''<partinfo>J23109</partinfo> Constitutive promoter family member (low expression)''' <br> pLow is a Low consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo><br> | + | '''<partinfo>J23109</partinfo> Constitutive promoter family member (low expression)''' <br> pLow is a Low consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. <partinfo>pSB3K3</partinfo>, <partinfo>BBa_J61002</partinfo> and <partinfo>pSB1AC3</partinfo><br> |
| - | '''<partinfo>R0010</partinfo> Promoter (lacI regulated)''' <br> This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds the CAP protein, which is generally present in E.coli and is asocciated with cell health and availability of glucose. The second binds LacI protein. <br> | + | '''<partinfo>R0010</partinfo> Promoter (lacI regulated)''' <br> This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds the CAP protein, which is generally present in E.coli and is asocciated with cell health and availability of glucose. The second binds LacI protein. <br> |
| - | '''<partinfo>I0500</partinfo> pBad/araC''' <br> This part is <br> | + | '''<partinfo>I0500</partinfo> pBad/araC''' <br> This part is <br> |
===Vectors=== | ===Vectors=== | ||
| - | '''<partinfo>pSB1AC3</partinfo> High copy BioBrick assembly plasmid''' | + | '''<partinfo>pSB1AC3</partinfo> High copy BioBrick assembly plasmid''' |
| - | + | ||
{| | {| | ||
|width='10%'| | |width='10%'| | ||
| Line 60: | Line 52: | ||
<I> iGEM Groningen 2009 </I> | <I> iGEM Groningen 2009 </I> | ||
|width='60%' valign='top'| | |width='60%' valign='top'| | ||
| - | The transformations with pSB1AC3 (containing different biobricks of own design) into ''E. coli'' TOP10 cells, growth on both antibiotics, and gel analysis (undigested and digested with the EcoRI and PstI) worked as expected. The high (copy) number of plasmids per cell make it an easy to work with plasmid, ideal for cloning and assembly work. | + | pSB1AC3 is a high copy number plasmid carrying ampicillin and chloramphenicol resistance. Together with pSB1A2 the vector was used for most assemblies of our team, because the high copy number made it easy to work with and easy isolation. The transformations with pSB1AC3 (containing different biobricks of own design) into ''E. coli'' TOP10 cells, growth on both antibiotics, and gel analysis (undigested and digested with the EcoRI and PstI) worked as expected. The high (copy) number of plasmids per cell make it an easy to work with plasmid, ideal for cloning and assembly work. |
|} | |} | ||
| - | + | '''<partinfo>pSB1A2</partinfo> pSB1A2 (Replaced by pSB1A3 in registry) ''' <br> pSB1A2 is a high copy number plasmid carrying ampicillin resistance. Together with pSB1AC3 the vector was used for most assemblies of our team, because the high copy number made it easy to work with and easy isolation. <br> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | '''<partinfo>pSB1A2</partinfo> pSB1A2 (Replaced by pSB1A3 in registry) ''' <br> pSB1A2 is a high copy number plasmid carrying ampicillin resistance. Together with pSB1AC3 the vector was used for most assemblies of our team, because the high copy number made it easy to work with and easy isolation. <br> | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | '''<partinfo>pSB2K3</partinfo> A low copy number base vector''' <br> A low copy number base vector which as a resistance against Kanamycin <br> | |
| - | '''<partinfo> | + | '''<partinfo>pSB3K3</partinfo> A low copy number base vector''' <br> A low copy number base vector which as a resistance against Kanamycin <br> |
| - | + | '''<partinfo>BBa_J61002</partinfo> A normal base vector''' <br> A normal base vector which has a rsistance against Ampicillin <br> | |
| - | '''<partinfo>BBa_J61035</partinfo> Vector we get with GVP''' <br> It's the backbone of our GVP part is and it a resistance against Ampicillin/Gentamycin<br> | + | '''<partinfo>BBa_J61035</partinfo> Vector we get with GVP''' <br> It's the backbone of our GVP part is and it a resistance against Ampicillin/Gentamycin<br> |
Revision as of 17:18, 17 October 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
|
|---|
Used Parts
TODOMaybe some general information of parts used from the registry.
- To get to the partsregistry site of a particular part one can click on the name of the part
- To see what our main findings where (no details or derivations) regarding a particular part one can click on more information
Miscellaneous
RBS ((Elowitz 1999) -- defines RBS efficiency)
|
iGEM Groningen 2009 |
The ligation of a part behind the RBS succeded, and was confirmed by gel (correct vector size) and sequencing with VF2. We used this part in combination with several genes for building our biobricks e.g. . |
Double terminator (B0012-B0011)
Double terminator consisting of BBa_B0012 and BBa_B0011
GVP Gas Vesicle Proteins
This parts creates gas vesicles inside the cell wich enables it to float. It consist of 6064 base pairs. It's backbone is
ccdB cell death gene
|
iGEM Groningen 2009 |
P1010 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or religated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result. The ccdB cell death gene worked as expected killing our E. coli TOP10 cells, and keeping our E. coli DB3 cells alive. After noticing the inconsistent sequencing result for the pSB2K3 plasmid with ccdB cell death gene, we decided to choose a different pSB2K3 plasmid with random part to continue with our assemblies. This to minimize the chance of unwanted surprises in the end. |
Promotors
Constitutive promoter family member (high expression)
pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. , and
Constitutive promoter family member (high expression, reference)
pHigh is a high consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. , and
Constitutive promoter family member (medium expression)
pMed is a medium consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. , and
Constitutive promoter family member (low expression)
pLow is a Low consituative promotor. It consists of about 35 base pairs. It has been placed in the following vectors. , and
Promoter (lacI regulated)
This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds the CAP protein, which is generally present in E.coli and is asocciated with cell health and availability of glucose. The second binds LacI protein.
pBad/araC
This part is
Vectors
High copy BioBrick assembly plasmid
|
iGEM Groningen 2009 |
pSB1AC3 is a high copy number plasmid carrying ampicillin and chloramphenicol resistance. Together with pSB1A2 the vector was used for most assemblies of our team, because the high copy number made it easy to work with and easy isolation. The transformations with pSB1AC3 (containing different biobricks of own design) into E. coli TOP10 cells, growth on both antibiotics, and gel analysis (undigested and digested with the EcoRI and PstI) worked as expected. The high (copy) number of plasmids per cell make it an easy to work with plasmid, ideal for cloning and assembly work. |
pSB1A2 (Replaced by pSB1A3 in registry)
pSB1A2 is a high copy number plasmid carrying ampicillin resistance. Together with pSB1AC3 the vector was used for most assemblies of our team, because the high copy number made it easy to work with and easy isolation.
A low copy number base vector
A low copy number base vector which as a resistance against Kanamycin
A low copy number base vector
A low copy number base vector which as a resistance against Kanamycin
A normal base vector
A normal base vector which has a rsistance against Ampicillin
It's the backbone of our GVP part is and it a resistance against Ampicillin/Gentamycin
