Part:BBa_I742103

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