Difference between revisions of "Part:BBa K777125"
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[[Image:Table_K777125-K777132.jpg|thumb|300px|right|'''Fig. 1:''' Overview of our related BioBricks with Anderson promoters and RFP in pSB1C3. (Activity according to Berkeley 2006)]] | [[Image:Table_K777125-K777132.jpg|thumb|300px|right|'''Fig. 1:''' Overview of our related BioBricks with Anderson promoters and RFP in pSB1C3. (Activity according to Berkeley 2006)]] | ||
− | This part is a | + | This part is a twin of an Anderson promoter in [https://parts.igem.org/Part:BBa_K777133 K777133], a pSB1C3 derived backbone. |
===Usage and Biology=== | ===Usage and Biology=== | ||
− | This construct can be useful in several ways, just like the | + | This construct can be useful in several ways, just like the similar part[https://parts.igem.org/Part:BBa_J61002 J61002]. However, if you plan to send coding or composite parts to the registry, they are already in the correct backbone here, [https://parts.igem.org/Part:pSB1C3 pSB1C3]. |
# You may use this plasmid to insert a promoter of your choice between the XbaI/EcoRI and SpeI sites. This will provide you with a downstream mRFP as a reporter. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here]. | # You may use this plasmid to insert a promoter of your choice between the XbaI/EcoRI and SpeI sites. This will provide you with a downstream mRFP as a reporter. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here]. | ||
# Furthermore, you can clone a gene of interest downstream of the constitutive Anderson promoters to express it at a desired rate. For this, the SpeI and PstI sites can be used. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here]. | # Furthermore, you can clone a gene of interest downstream of the constitutive Anderson promoters to express it at a desired rate. For this, the SpeI and PstI sites can be used. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here]. | ||
# It can provide a nice selection system for inserting your basic parts into the pSB1C3 vector via the EcoRI/XbaI and PstI site. If you use a construct containing a strong promoter e.g. K777125, colonies containing a self-ligated plasmid after transformation will turn red and you can select for the non-red colonies. | # It can provide a nice selection system for inserting your basic parts into the pSB1C3 vector via the EcoRI/XbaI and PstI site. If you use a construct containing a strong promoter e.g. K777125, colonies containing a self-ligated plasmid after transformation will turn red and you can select for the non-red colonies. | ||
<br> | <br> | ||
− | * Here is the complete sequence as an [https://static.igem.org/mediawiki/ | + | We also used this construct as a control for our [https://parts.igem.org/Image:YhjH_motility.jpg swimming assays]. |
− | <br> | + | <br><br> |
+ | * Here is the complete sequence as an [https://static.igem.org/mediawiki/2012/0/07/K777125-K777132.zip ApE file] | ||
+ | <br><br> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 15:48, 28 September 2012
Constitutive J23100 promoter
This part is a twin of an Anderson promoter in K777133, a pSB1C3 derived backbone.
Usage and Biology
This construct can be useful in several ways, just like the similar partJ61002. However, if you plan to send coding or composite parts to the registry, they are already in the correct backbone here, pSB1C3.
- You may use this plasmid to insert a promoter of your choice between the XbaI/EcoRI and SpeI sites. This will provide you with a downstream mRFP as a reporter. Look up the specifications for restriction sites here.
- Furthermore, you can clone a gene of interest downstream of the constitutive Anderson promoters to express it at a desired rate. For this, the SpeI and PstI sites can be used. Look up the specifications for restriction sites here.
- It can provide a nice selection system for inserting your basic parts into the pSB1C3 vector via the EcoRI/XbaI and PstI site. If you use a construct containing a strong promoter e.g. K777125, colonies containing a self-ligated plasmid after transformation will turn red and you can select for the non-red colonies.
We also used this construct as a control for our swimming assays.
- Here is the complete sequence as an ApE file
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 8
Illegal NheI site found at 31 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]