Plasmid_Backbone

Part:pSB3C01

Designed by: Vinoo Selvarajah   Group: Registry   (2019-04-04)
Revision as of 10:10, 20 October 2020 by Nuria GA (Talk | contribs) (Uppsala 2020's Improvement)


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


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2713
    Illegal PstI site found at 12
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2713
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2713
    Illegal PstI site found at 12
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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 pEven1 plasmid pSB3C11 (BBa_K3425001). This new plasmid was used for various clonings and it works as expected.

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