Difference between revisions of "Part:pSB6A1"
Line 1: Line 1:
 
__NOTOC__
 
__NOTOC__
−
<partinfo>BBa_I739201 short</partinfo>
+
<partinfo>pSB6A1 short</partinfo>
  
 
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>
Line 6: Line 6:
  
 
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".
−
<br><br>
+
<br>
 
===Purpose===
 
===Purpose===
−
<p>This plasmid was designed for the [http://parts.mit.edu/igem07/index.php/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://parts.mit.edu/igem07/index.php?title=ETHZ/Biology/Lab here].
+
<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>
  
Line 16: Line 16:
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
−
<partinfo>BBa_I739201 SequenceAndFeatures</partinfo>
+
<partinfo>pSB6A1 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
−
<partinfo>BBa_I739201 parameters</partinfo>
+
<partinfo>pSB6A1 parameters</partinfo>
 
<!-- -->
 
<!-- -->

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.

Fig. 1: Map of the pBR322BB1 plasmid
Fig. 2: Map of the original pBR322 plasmid

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


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 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4001
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4001
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 3075
    Illegal SapI site found at 1992