Difference between revisions of "Part:BBa I739203"
 
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__NOTOC__
 
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<partinfo>BBa_I739202 short</partinfo>
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<partinfo>BBa_I739203 short</partinfo>
  
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Plasmid pCK01MCS1 is based on pCK01 [1], a low-copy cloning vector that confers chloramphenicol resistance (CmR) and harbours an oriV.<br>
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Plasmid pBR322BB2 (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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[[Image:Pck01 Andy.jpg|350px]]
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[[Image:MapI739203.jpg|left|thumb|'''Fig. 1:''' Map of the pBR322BB2 plasmid|250px]][[Image:Mappbr322.jpg|right|thumb|'''Fig. 2:''' Map of the original pBR322 plasmid|250px]]
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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], a MCS harbouring a SpeI site was introduced by PCR. This MCS is the only difference between pCKMCS1 and pCK01:<br>
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[[Image:PCK01-2.jpg]]
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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 PstI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original ampicillin resistance and only tetracyclin resistance is left. The term "BB2" in pBR322BB2 refers to "BioBrick version 2".
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<br>
 
===Purpose===
 
===Purpose===
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<p>This plasmid was designed for the [http://parts.mit.edu/igem07/index.php/ETHZ ETHZ iGEM 2007 project] and contains... </p>
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<p>This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and was used as a backup plasmid for [https://parts.igem.org/wiki/index.php?title=Part:BBa_I739201 pBR322BB1]. More information on the plasmid and the used assembly strategy can be found [http://2007.igem.org?title=ETHZ/Biology/Lab here].
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</p><br>
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=== References ===
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<p>
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[1] Fernández S, de Lorenzo V, Pérez-Martín J <i>"Activation of the transcriptional regulator XylR of Pseudomonas putida by release of repression between functional domains"</i>, Mol Microbiol. 16(2):205-13, 2000</p>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_I739203 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_I739202 parameters</partinfo>
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<partinfo>BBa_I739203 parameters</partinfo>
 
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Latest revision as of 08:38, 25 October 2007

pBR322BB2

Plasmid pBR322BB2 (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 pBR322BB2 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 PstI restriction sites and an appropriate multiple cloning site (MCS) was introduced in this position. However, this leads to a loss of the original ampicillin resistance and only tetracyclin resistance is left. The term "BB2" in pBR322BB2 refers to "BioBrick version 2".

Purpose

This plasmid was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and was used as a backup plasmid for pBR322BB1. 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
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3638
    Illegal SpeI site found at 3621
    Illegal PstI site found at 3607
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 229
    Illegal SpeI site found at 3621
    Illegal PstI site found at 3607
    Illegal NotI site found at 3612
    Illegal NotI site found at 3645
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 375
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3638
    Illegal SpeI site found at 3621
    Illegal PstI site found at 3607
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3638
    Illegal SpeI site found at 3621
    Illegal PstI site found at 3607
    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