Difference between revisions of "Part:BBa I751000"

Latest revision as of 03:03, 27 October 2007

pBR322 (Pst I site mutated)

pBR322 has Pst I site in its Amp resistant gene.

This Pst site can be digested at the same time BioBrick parts are inserted even into other regions, which inhibits convenient gene construction.

Therefore, by replacing A at the 546th with C (A546C), we mutated the Pst site not to be digested by Pst. Introducing this mutation allowed us to aim at constructing pBR standard plasmid.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 229
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 375
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 401
Illegal NgoMIV site found at 769
Illegal NgoMIV site found at 929
Illegal NgoMIV site found at 1283
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 3433
Illegal SapI site found at 2350

@@ Line 2: / Line 2: @@
 <partinfo>BBa_I751000 short</partinfo>
-pBR322 has Pst I site in its Amp resistant gene. It prevents us from introducing Biobrick site(Pst I included) to this plasmid.
+pBR322 has Pst I site in its Amp resistant gene.
-For that reason, we introduced a silent mutation at the site, in order not to restrict Amp resistant gene. The mutation is A546C(DNA sequence) in the Amp resistant gene. Introducing this mutation allowed us to aim at constructing pBR standard plasmid.
+This Pst site can be digested at the same time BioBrick parts are inserted even into other regions, which inhibits convenient gene construction.
+Therefore, by replacing A at the 546th with C (A546C), we mutated the Pst site not to be digested by Pst.
+Introducing this mutation allowed us to aim at constructing pBR standard plasmid.
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