Difference between revisions of "Part:BBa K590011"
(→Usage and Biology) |
|||
| Line 7: | Line 7: | ||
This is a high copy plasmid backbone which has Chloramphenicol resistance, with pLac promoter and GFP. This is a high efficiency Gibson Assembly vector. In a pGA vector evaluation, we determined the cloning efficiency by by dividing the # of bright colonies by the (# of total colonies - # of background colonies). The background colonies were determined by a control sample containing 100 picograms of just the pGA backbone | This is a high copy plasmid backbone which has Chloramphenicol resistance, with pLac promoter and GFP. This is a high efficiency Gibson Assembly vector. In a pGA vector evaluation, we determined the cloning efficiency by by dividing the # of bright colonies by the (# of total colonies - # of background colonies). The background colonies were determined by a control sample containing 100 picograms of just the pGA backbone | ||
| − | On a average of 6 plates, the Gibson efficiency was determined to be 0.991129032 ~ '''99%!'''. | + | ===Characterization=== |
| − | + | A sister plasmid ([https://parts.igem.org/Part:BBa_K590010 pGA1A3]) with the same BglBrick prefix and suffix regions has been shown to have much higher efficiency than the equivalent pSB vector. On a average of 6 plates, the Gibson efficiency was determined to be 0.991129032 ~ '''99%!'''. See [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonResults#Comparison_between_pGA_and_pSB_vectors Gibson Assembly efficiency assay] page for details on the protocol and efficiency measurements. | |
| − | + | <center>https://static.igem.org/mediawiki/2011/d/d1/Washington_pGAefficiency_summary.jpg</center> | |
<!-- --> | <!-- --> | ||
Revision as of 23:17, 28 September 2011
pGA1C3, Gibson assembly plasmid (bglBrick) with pLac-GFP insert
This Gibson Cloning friendly 1C3 plasmid backbone was made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] as part of the [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonVectors Gibson Assembly toolkit].
Usage and Biology
This is a high copy plasmid backbone which has Chloramphenicol resistance, with pLac promoter and GFP. This is a high efficiency Gibson Assembly vector. In a pGA vector evaluation, we determined the cloning efficiency by by dividing the # of bright colonies by the (# of total colonies - # of background colonies). The background colonies were determined by a control sample containing 100 picograms of just the pGA backbone
Characterization
A sister plasmid (pGA1A3) with the same BglBrick prefix and suffix regions has been shown to have much higher efficiency than the equivalent pSB vector. On a average of 6 plates, the Gibson efficiency was determined to be 0.991129032 ~ 99%!. See [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonResults#Comparison_between_pGA_and_pSB_vectors Gibson Assembly efficiency assay] page for details on the protocol and efficiency measurements.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2051 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2051 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2051
Illegal BglII site found at 2066
Illegal BamHI site found at 1
Illegal XhoI site found at 16
Illegal XhoI site found at 1035
Illegal XhoI site found at 1927 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2051 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2051 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.