Difference between revisions of "Part:pSB3C01"
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==Uppsala 2020's Improvement== | ==Uppsala 2020's Improvement== | ||
| + | <i><b>TEAMNAME</b> UofUppsala <b>YEAR</b> 2020 <b>Authors</b> Nuria Garriga Alonso, Bjorn Ancker Persson, Tereza Hubackova, Dorottya Marko, Hui Yu </i> | ||
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The illegal BsaI site was removed by PCR mutagenesis to obtain the new pEven1 plasmid pSB3C11 ([[Part:BBa_K3425001|<partinfo>BBa_K3425001</partinfo>]]). This new plasmid was used for various clonings and it works as expected. | The illegal BsaI site was removed by PCR mutagenesis to obtain the new pEven1 plasmid pSB3C11 ([[Part:BBa_K3425001|<partinfo>BBa_K3425001</partinfo>]]). This new plasmid was used for various clonings and it works as expected. | ||
Moreover, pSB3C01 was also used for cloning and it also works as expected. However, we recommend using the new plasmids without an illegal BsaI site in order to avoid compromising the efficiency. | Moreover, pSB3C01 was also used for cloning and it also works as expected. However, we recommend using the new plasmids without an illegal BsaI site in order to avoid compromising the efficiency. | ||
Latest revision as of 10:15, 22 October 2020
pEven1 Loop Vector based on pSB3C5
pEven1 Loop Vector based on pSB3C5
iGEM Type IIS Vectors
pEven Loop Vectors (for Level 2)
When four Level 1 parts (TUs) are assembled into a Multi-Transcriptional Unit (MTU) into the following plasmid backbones (with BBa_J04455), the MTU will be flanked by BsaI restriction sites and these 4 bp Fusion Sites.
| Registry Name | Loop Name | Fusion Site 5' | MTU | Fusion Site 3' |
|---|---|---|---|---|
| pSB3C01 | pEven1 | GGAG | MTU 1 | TACT |
| pSB3C02 | pEven2 | TACT | MTU 2 | AATG |
| pSB3C03 | pEven3 | AATG | MTU 3 | GCTT |
| pSB3C04 | pEven4 | GCTT | MTU 4 | CGCT |
Note:
- The documented sequence shows an additional BsaI site. This would impact assembly out of (but not into) this set of vectors. A new set of vectors should be created or used in the long term.
- Currently, Registry samples of these plasmid backbones have not been fully sequenced. Only their prefix and suffix along with their insert BBa_J04455 have been verified.
The iGEM Type IIS assembly standard is based on MoClo and Loop
- A Modular Cloning System for Standardized Assembly of Multigene Constructs
- Loop assembly: a simple and open system for recursive fabrication of DNA circuits
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2713
Illegal PstI site found at 12 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2713
Illegal NheI site found at 1399
Illegal PstI site found at 12 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2713 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2713
Illegal PstI site found at 12 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2713
Illegal PstI site found at 12
Illegal AgeI site found at 990
Illegal AgeI site found at 1313 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 285
Illegal BsaI site found at 2719
Illegal BsaI.rc site found at 6
Uppsala 2020's Improvement
TEAMNAME UofUppsala YEAR 2020 Authors Nuria Garriga Alonso, Bjorn Ancker Persson, Tereza Hubackova, Dorottya Marko, Hui Yu
The illegal BsaI site was removed by PCR mutagenesis to obtain the new pEven1 plasmid pSB3C11 (BBa_K3425001). This new plasmid was used for various clonings and it works as expected.
Moreover, pSB3C01 was also used for cloning and it also works as expected. However, we recommend using the new plasmids without an illegal BsaI site in order to avoid compromising the efficiency.