Difference between revisions of "Part:pSB4A3"

 
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pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream.  A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region.
 
pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream.  A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region.
  
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===Usage and Biology===
 
===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>pSB4A3 SequenceAndFeatures</partinfo>
 
<partinfo>pSB4A3 SequenceAndFeatures</partinfo>
  
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>pSB4A3 parameters</partinfo>
 
<partinfo>pSB4A3 parameters</partinfo>
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Revision as of 02:17, 5 June 2006


pSB4A3 (Replaced by pSB4A5)

pSB4A3-1 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream. A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region.

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 3318
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3324
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3318
    Illegal XhoI site found at 177
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3318
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3318
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3333
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 2360