Difference between revisions of "Part:BBa I742103"

Revision as of 17:07, 25 October 2007

pTG262 Plasmid (Gram negative to Gram positive shuttle vector)

This plasmid contains an origin of replication which is capable of replicating in E. coli and several gram positive organisms including Lactobacillus, Lactococcus and Bacillus. The plasmid has chloramphenicol, neomycin & gentamicin resistance genes. It also has EcoRI and PstI sites 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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 <partinfo>BBa_I742103 short</partinfo>
-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.
+This plasmid contains an origin of replication which is capable of replicating in ''E. coli'' and several gram positive organisms including ''Lactobacillus'', ''Lactococcus'' and ''Bacillus''.  The plasmid has chloramphenicol, neomycin & gentamicin resistance genes.  It also has EcoRI and PstI sites 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).