Difference between revisions of "Part:pSB1C3:Design"

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<partinfo>pSB1C3 SequenceAndFeatures</partinfo>
 
<partinfo>pSB1C3 SequenceAndFeatures</partinfo>
  
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===Original Sequence===
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In Spring 2011, pSB1C3 was sequenced using several primers. The current sequence reflects the most recent sequencing of pSB1C3 (w/ BBa_J04450, located in SP 4000 Well 2A). However for posterity, we have included the old sequence that was originally specified in the Registry ([https://parts.igem.org/Image:PSB1C3-Text-Archival-5-4-11.txt PSB1C3-Text-Archival-5-4-11.txt]) as well as images of the original sequence with markers pointing out the changes made:
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[[Image:PSB1C3-Seq-Archival-5-4-11.png|300px]]
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[[Image:PSB1C3-Seq-Archival-Text.png|300px]]
  
 
===Design Notes===
 
===Design Notes===
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====Freiburg's pSB1C3 varient of 2010====
 
====Freiburg's pSB1C3 varient of 2010====
 
The iGEM team Freiburg created a modified version of the pSB1C3 in which the two restriction sites SspI and PvuII were removed via site directed mutagenesis. These restriction sites and two others that are not present in the pSB1C3 (BamHI and SalI) were used as single cutting restriction sites to replace the loop coding sequences of the Adeno-associated Virus. For this purpose scar-less cloning is necessary, because of the unpredictable consequences on the viral vector performance arising from mutations or insertions. The final capsid gene adheres fully to the RFC_10 standard.<br>
 
The iGEM team Freiburg created a modified version of the pSB1C3 in which the two restriction sites SspI and PvuII were removed via site directed mutagenesis. These restriction sites and two others that are not present in the pSB1C3 (BamHI and SalI) were used as single cutting restriction sites to replace the loop coding sequences of the Adeno-associated Virus. For this purpose scar-less cloning is necessary, because of the unpredictable consequences on the viral vector performance arising from mutations or insertions. The final capsid gene adheres fully to the RFC_10 standard.<br>
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The sequence and a more detailled description can be found under the BioBrick ID BBa_K404200.
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The sequence and a more detailled description can be found under the BioBrick ID [https://parts.igem.org/Part:BBa_K404200 BBa_K404200].
  
 
===Source===
 
===Source===

Latest revision as of 17:39, 5 May 2011

High copy BioBrick assembly plasmid


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 2049
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2055
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2049
    Illegal XhoI site found at 1033
    Illegal XhoI site found at 1925
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 2049
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 2049
    Plasmid lacks a suffix.
    Illegal XbaI site found at 2064
    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.

Original Sequence

In Spring 2011, pSB1C3 was sequenced using several primers. The current sequence reflects the most recent sequencing of pSB1C3 (w/ BBa_J04450, located in SP 4000 Well 2A). However for posterity, we have included the old sequence that was originally specified in the Registry (PSB1C3-Text-Archival-5-4-11.txt) as well as images of the original sequence with markers pointing out the changes made:

PSB1C3-Seq-Archival-5-4-11.png PSB1C3-Seq-Archival-Text.png

Design Notes

Eliminated ampicillin resistance.

Subversions

Freiburg's pSB1C3 varient of 2010

The iGEM team Freiburg created a modified version of the pSB1C3 in which the two restriction sites SspI and PvuII were removed via site directed mutagenesis. These restriction sites and two others that are not present in the pSB1C3 (BamHI and SalI) were used as single cutting restriction sites to replace the loop coding sequences of the Adeno-associated Virus. For this purpose scar-less cloning is necessary, because of the unpredictable consequences on the viral vector performance arising from mutations or insertions. The final capsid gene adheres fully to the RFC_10 standard.
The sequence and a more detailled description can be found under the BioBrick ID BBa_K404200.

Source

This part was derived from pSB1AC3-P1010.

References