Plasmid
pTG262

Part:BBa_I742103

Designed by: sarah hollingshead   Group: iGEM07_Edinburgh   (2007-10-06)
Revision as of 12:40, 23 October 2007 by Cfrench (Talk | contribs)

pTG262 Plasmid (Gram negative to Gram positive shuttle vector)

Plasmid contains an origin of replication which is capable of replicating in E. coli and several gram positive organisms including Lactobacillus, Lactococcus and Bacillus. Plasmid has chloramphenicol, neomycin & gentamicin resistance genes. It also has a EcoRI and PstI site with an XbaI site situated in the middle so biobricks can be inserted. Insertion of any biobrick between the unique EcoRI and PstI sites removes the only XbaI site, and there are no SpeI sites, so the product is a compliant biobrick. pTG262 was generously provided by Dr. Mike Gasson of the Institute of Food Research, Norwich, UK. Recommended selection with chloramphenicol is as follows: E. coli 15 mg/l; Bacillus subtilis 10 mg/l; Lactococcus lactis 5 mg/l; Lactobacillus spp. 7.5 mg/l.

Gene instability may occur with certain inserts (we have not experienced this so far with the biobricks we have inserted).

For more information about pTG262 visit the [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Gene_Expression_In_Lactobacillus Edinburgh Yoghurt Wiki]


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 463
    Illegal NheI site found at 5313
    Illegal PstI site found at 424
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 463
    Illegal BglII site found at 2252
    Illegal BamHI site found at 442
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
    Illegal AgeI site found at 2641
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2427


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