Difference between revisions of "Part:pSB3C01"
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+ | ==Uppsala 2020's Improvement== | ||
+ | The illegal BsaI site was removed by PCR mutagenesis to obtain the new pEven plasmid pSB3C11 ([[Part:BBa_K3425001|<partinfo>BBa_K3425001</partinfo>]]). This new plasmid was used for various clonings and it works as expected. |
Revision as of 10:08, 20 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
Assembly Compatibility:
- 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
The illegal BsaI site was removed by PCR mutagenesis to obtain the new pEven plasmid pSB3C11 (BBa_K3425001). This new plasmid was used for various clonings and it works as expected.