Part:BBa_K2656200

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 composite parts from the [http://2018.igem.org/Team:Valencia_UPV/Part_Collection UPV iGEM 2018 Printeria Part Collection] into the BioBrick standard.

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.