Part:BBa_K1033206

Lactobacillus shuttle vector pSBLbC

The backbone pSBLbC is a shuttle vector between E. coli, Lacotbacillus, and some other lactic acid bacteria. It has been used for subcloning in E. coli and to express a chromoprotein amilCP. For use in Lactobacillus we recommend the version with erythromycin resistance (pSBLbE) since that has been successfully used to transform Lactobacillus.

Fig 1: E. coli D5-alpha with pSBLbC to which amilCP and lactobacillus promotor CP29 have been subcloned

It was made by replacing the replicon of BioBrick compatible plasmid pSB4C15 (BBa_K864001) with a broad range replicon from the engineered plasmid pJP059. The replicon is also referred to as pSH71 and is related to pWV01 and that family of rolling circle replicating plasmids.

We also had to change the promotor of the chloramphenicol resistance cassette to one that would initiate transcription effectively in Lactobacillus we tried constitutive promotors but finally got CP29 to work.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 3036
  Illegal suffix found in sequence at 1
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3036
  Illegal NheI site found at 1981
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 3042
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3036
  Illegal BamHI site found at 1960
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 3036
  Illegal suffix found in sequence at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 3036
  Illegal XbaI site found at 3051
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  COMPATIBLE WITH RFC[1000]