Part:BBa_K128008

pJK650 plasmid

Expression vector pJK650 from L. delbruckii bacteria. Easiest if the plasmid is grown up in Lactococcus lactis and extracted from there. Confers Erythromycin resistance; select in Lactococcus lactis with 5 ug/ml Em, select in Lactobacillus with 7.5 ug/ml Em.

With electroporation procedures reported by Serror et al. (App. and Env. Microbiology 2002), pJK650 is compatible with Lactobacillus delbruckii subsp. bulgaricus strains VI104 and ATCC 11842 and Lactobacillus delbruckii subsp. lactis strain ATCC 10697. Transformation efficiency ~ 10^2 transformants/ug.

For detailed electroporation protocols, please visit the MIT 2008 iGEM wiki or the Lactobacillus protocols section on openwetware.org

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 7384
  Illegal XbaI site found at 7337
  Illegal XbaI site found at 7388
  Illegal SpeI site found at 944
  Illegal PstI site found at 7348
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 7384
  Illegal NheI site found at 1541
  Illegal SpeI site found at 944
  Illegal PstI site found at 7348
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 7384
  Illegal BglII site found at 1554
  Illegal BglII site found at 3036
  Illegal BglII site found at 5286
  Illegal BglII site found at 5442
  Illegal BglII site found at 6134
  Illegal BamHI site found at 7353
  Illegal XhoI site found at 155
  Illegal XhoI site found at 1502
  Illegal XhoI site found at 2621
  Illegal XhoI site found at 2714
  Illegal XhoI site found at 4370
  Illegal XhoI site found at 4769
  Illegal XhoI site found at 5043
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 7384
  Illegal XbaI site found at 7337
  Illegal XbaI site found at 7388
  Illegal SpeI site found at 944
  Illegal PstI site found at 7348
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 7384
  Illegal XbaI site found at 7337
  Illegal XbaI site found at 7388
  Illegal SpeI site found at 944
  Illegal PstI site found at 7348
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 5956
  Illegal BsaI.rc site found at 2181
  Illegal BsaI.rc site found at 2866
  Illegal BsaI.rc site found at 6796
  Illegal SapI.rc site found at 4912
  

Restriction Site Map of pJK650: