Plasmid_Backbone

Part:BBa_K2656200

Designed by: Carolina Ropero   Group: iGEM18_Valencia_UPV   (2018-09-21)
Revision as of 21:56, 17 October 2018 by Carolinarp (Talk | contribs)


pBIOBRICKATOR

BBa_K2656200 (pBioBrickator backbone) is a modification of the pSB1C3 backbone to receive transcriptional units previously assembled in a [http://2018.igem.org/Team:Valencia_UPV/Design#Level1 GB alpha 1] vector.

To do so, it includes two divergently orientated BsmBI Type IIS recognition sites, whose cleavage leaves equivalent overhangs to those of the alpha 1 BsmBI digestion (Figure 1). Thus, it can be used to ligate any transcriptional unit assembled in an alpha 1 GB vector into a BioBrick compatible vector, so adapting the composite part into the BB grammar with an easy GB one-pot reaction.

This backbone has been used to adapt each our our composite parts of the [http://2018.igem.org/Team:Valencia_UPV/Part_Collection UPV iGEM 2018 Printeria Part Collection] into the BioBrick standards.

domestication.
Figure 1. Transcriptional unit assembled in an alpha 1 GoldenBraid vector (KmR). Digestion with BsmBI Type IIS endonuclease leaves complementary overhangs (5'-GGAG-3' and 3'-CATG-5') to the pBioBrickator vector digest.










Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 5
    Plasmid lacks a suffix.
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5
    Illegal NheI site found at 44
    Illegal NheI site found at 67
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
    Illegal NotI site found at 11
    Illegal NotI site found at 928
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5
    Illegal XhoI site found at 1952
    Illegal XhoI site found at 2844
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5
    Illegal suffix found at 921
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5
    Plasmid lacks a suffix.
    Illegal XbaI site found at 20
    Illegal SpeI site found at 921
    Illegal PstI site found at 935
    Illegal AgeI site found at 653
    Illegal AgeI site found at 765
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


[edit]
Categories
Parameters
None