Difference between revisions of "Part:BBa I742123"

Revision as of 17:52, 25 October 2007

Multi-host vector pTG262 converted to BioBrick vector

This plasmid was derived from pTG262 by insertion of a biobrick encoding Red Fluorescent Protein between the unique EcoRI and PstI restriction sites. This eliminated the only native XbaI site present in the plasmid, and since there are no native SpeI sites, the resulting plasmid is a compliant BioBrick vector. New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies.The parent plasmid pTG262 has a multi host replication origin and replicates in a range of gram positive and gram negative bacteria including E. coli, Bacillus subtilis, and Lactobacillus and Lactococcus spp. We hope that it will also replicate in other Gram negative bacteria. At the time of writing we have successfully transformed E. coli and B. subtilis with this construct, selecting with chloramphenicol (15 and 10 mg/l respectively). However, RFP did not seem to be expressed in B. subtilis; possibly a promoter issue.

More information about pTG262 may be found on the [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Gene_Expression_In_Lactobacillus Edinburgh Yoghurt Wiki]

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 5543
Illegal NheI site found at 694
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 5549
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 5543
Illegal BglII site found at 3754
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found at 5543
Illegal suffix found at 2
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found at 5543
Plasmid lacks a suffix.
Illegal XbaI site found at 5558
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal AgeI site found at 3365
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal SapI.rc site found at 3578

@@ Line 2: / Line 2: @@
 <partinfo>BBa_I742123 short</partinfo>
-biobricked version of pTG262
+This plasmid was derived from pTG262 by insertion of a biobrick encoding Red Fluorescent Protein between the unique EcoRI and PstI restriction sites. This eliminated the only native XbaI site present in the plasmid, and since there are no native SpeI sites, the resulting plasmid is a compliant BioBrick vector. New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies.The parent plasmid pTG262 has a multi host replication origin and replicates in a range of gram positive and gram negative bacteria including ''E. coli'', ''Bacillus subtilis'', and ''Lactobacillus'' and ''Lactococcus'' spp. We hope that it will also replicate in other Gram negative bacteria. At the time of writing we have successfully transformed ''E. coli'' and ''B. subtilis'' with this construct, selecting with chloramphenicol (15 and 10 mg/l respectively).  However, RFP did not seem to be expressed in ''B. subtilis''; possibly a promoter issue.
-Plasmid has a multi host replication origin and is competant in range of gram positive and gram negative bacteria.
 More information about pTG262 may be found on the [http://parts.mit.edu/igem07/index.php/Edinburgh/Yoghurt/Design#Gene_Expression_In_Lactobacillus Edinburgh Yoghurt Wiki]