Plasmid_Backbone

Part:BBa_K3781103:Design

Designed by: Nicolas Bayer   Group: iGEM21_TU_Kaiserslautern   (2021-10-09)


pAGM1299


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2477
    Illegal XbaI site found at 2450
    Illegal PstI site found at 2438
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2477
    Illegal PstI site found at 2438
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2477
    Illegal BamHI site found at 2456
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2477
    Illegal XbaI site found at 2450
    Illegal PstI site found at 2438
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2477
    Illegal XbaI site found at 2450
    Illegal PstI site found at 2438
    Illegal NgoMIV site found at 1081
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2225
    Illegal BsaI.rc site found at 2835
    Illegal SapI site found at 4
    Illegal SapI.rc site found at 2162


Design Notes

This plasmid backbone was not designed by us, yet simply utilized from the Modular Cloning Toolbox established by Weber et al.[1]. We purchased it from our supervising working group that is working with MoClo in Chlamydomonas reinhardtii. Despite this part not being thought up, engineered or modified by us, we decided to include it into the registry in order to give possible users of our MocloMania collection a holistic view on the genetic material needed to build their own MoClo library and carry out their desired MoClo assemblies.


Source

The plasmid's sequence is directly derived from the creators of the Weber et al. Modular Cloning Kit. It can be found online as the Addgene plasmid named pAGM1299, #47988. The basic make-up of the plasmid is an alteration of the standard pUC19/pUC18 plasmids that have been a common lab staple for cloning in E. Coli for decades.[2] The sequences included within the plasmid, such as the N-terminal beta-galactosidase fragment lacZ-alpha and the ori are modifications of genes found within the genome of many E. Coli strains.


References

  1. ↑ Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S (2011) A Modular Cloning System for Standardized Assembly of Multigene Constructs. PLoS ONE 6(2): e16765. https://doi.org/10.1371/journal.pone.0016765
  2. ↑ C. Yanisch-Perron, J. Vieira, J. Messing: Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. In: Gene. Band 33, Nummer 1, 1985, S. 103–119. PMID 2985470.