Plasmid

Part:BBa_K128007

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

pLEM415 plasmid

Expression vector pLEM415 with biobrick sites inserted. A shuttle vector (derived from L. reuteri - E. coli) obtained by ligating PstI-digested pLEM5 with PstI-linearized E. coli vector pBII. Able to replicate in E. Coli and species of Lactobacillus. Confers Erythromycin resistance; select in E. coli with 150 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), pLEM415 is compatible with Lactobacillus bulgaricus strain VI104 and Lactobacillus delbruckii subsp. lactis strain ATCC 10697. Transformation efficiency ~ 10^3 - 10^4 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 701
    Illegal XbaI site found at 4102
    Illegal SpeI site found at 4096
    Illegal PstI site found at 707
    Illegal PstI site found at 4078
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 701
    Illegal NheI site found at 2678
    Illegal SpeI site found at 4096
    Illegal PstI site found at 707
    Illegal PstI site found at 4078
    Illegal NotI site found at 4108
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 701
    Illegal BglII site found at 2076
    Illegal BamHI site found at 4090
    Illegal XhoI site found at 668
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 701
    Illegal XbaI site found at 4102
    Illegal SpeI site found at 4096
    Illegal PstI site found at 707
    Illegal PstI site found at 4078
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 701
    Illegal XbaI site found at 4102
    Illegal SpeI site found at 4096
    Illegal PstI site found at 707
    Illegal PstI site found at 4078
    Illegal NgoMIV site found at 129
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5484
    Illegal EcoRI site found at 701
    Illegal XbaI site found at 4102
    Illegal SpeI site found at 4096
    Illegal PstI site found at 707
    Illegal PstI site found at 4078


Restriction Site Map of pLEM415: PLEM415 unique cut sites.jpg

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