Difference between revisions of "Part:pSB6A1"
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Plasmid pBR322BB1 (Fig. 1) is based on pBR322 (Fig. 2), a low-copy cloning vector (15-20 copies per cell) that confers ampicillin (ApR) and tetracylin (TcR) resistance and harbours the pMB1 origin of replication.<br> | Plasmid pBR322BB1 (Fig. 1) is based on pBR322 (Fig. 2), a low-copy cloning vector (15-20 copies per cell) that confers ampicillin (ApR) and tetracylin (TcR) resistance and harbours the pMB1 origin of replication.<br> | ||
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In order to make this plasmid compatible with the [https://parts.igem.org/wiki/index.php/Assembly:Standard_assembly BioBrick Standard assembly process], the original pBR322 was digested at its EcoRI and BamHI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original tetracyclin resistance and only ampicillin resistance is left. Since the original plasmid also contains a PstI restriction site in the bla gene, the GCA codon at this position was changed to GTA by site directed mutagenesis. The term "BB1" in pBR322BB1 refers to "BioBrick version 1". | In order to make this plasmid compatible with the [https://parts.igem.org/wiki/index.php/Assembly:Standard_assembly BioBrick Standard assembly process], the original pBR322 was digested at its EcoRI and BamHI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original tetracyclin resistance and only ampicillin resistance is left. Since the original plasmid also contains a PstI restriction site in the bla gene, the GCA codon at this position was changed to GTA by site directed mutagenesis. The term "BB1" in pBR322BB1 refers to "BioBrick version 1". | ||
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===Purpose=== | ===Purpose=== | ||
− | <p>This plasmid was designed for the [http:// | + | <p>This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and is used for the following BioBricks: [https://parts.igem.org/wiki/index.php/Part:BBa_I739001 ''Part 1''], [https://parts.igem.org/wiki/index.php/Part:I739002 ''Part 2''], [https://parts.igem.org/wiki/index.php/Part:BBa_I739003 ''Part 3''] and the composites [https://parts.igem.org/wiki/index.php/Part:BBa_I739012 BBa_I739012] and [https://parts.igem.org/wiki/index.php/Part:BBa_I739013 BBa_I739013]. More information on the plasmid and the used assembly strategy can be found [http://2007.igem.org?title=ETHZ/Biology/Lab here]. |
</p><br> | </p><br> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
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<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
− | <partinfo> | + | <partinfo>pSB6A1 parameters</partinfo> |
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Revision as of 08:41, 25 October 2007
pBR322 converted to BioBrick vector (formally BBa_I739201)
Plasmid pBR322BB1 (Fig. 1) is based on pBR322 (Fig. 2), a low-copy cloning vector (15-20 copies per cell) that confers ampicillin (ApR) and tetracylin (TcR) resistance and harbours the pMB1 origin of replication.
In order to make this plasmid compatible with the BioBrick Standard assembly process, the original pBR322 was digested at its EcoRI and BamHI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original tetracyclin resistance and only ampicillin resistance is left. Since the original plasmid also contains a PstI restriction site in the bla gene, the GCA codon at this position was changed to GTA by site directed mutagenesis. The term "BB1" in pBR322BB1 refers to "BioBrick version 1".
Purpose
This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and is used for the following BioBricks: Part 1, Part 2, Part 3 and the composites BBa_I739012 and BBa_I739013. More information on the plasmid and the used assembly strategy can be found [http://2007.igem.org?title=ETHZ/Biology/Lab here].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4001
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4007 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4001 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 4001
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 4001
Plasmid lacks a suffix.
Illegal XbaI site found at 4016
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 43
Illegal NgoMIV site found at 411
Illegal NgoMIV site found at 571
Illegal NgoMIV site found at 925 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 3075
Illegal SapI site found at 1992