Plasmid

Part:BBa_K128008

Designed by: Derek Ju   Group: iGEM08_MIT   (2008-10-29)

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 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


Restriction Site Map of pJK650:

PJK650 unique cut sites.jpg

[edit]
Categories
Parameters
None