Difference between revisions of "Part:BBa I50042:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_I50042 short</partinfo>
 
<partinfo>BBa_I50042 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The replication origin boundaries were chosen quite liberally (i.e. they might include more sequence than is strictly necessary) because a different version of this origin (BBa_I50040) was shown to be non-functional.
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The original design for [[Part:BBa_I50042|BBa_I50042]] was [[Part:BBa_I50040|BBa_I50040]]. Synthesis of [[Part:BBa_I50040|BBa_I50040]] (designed pSC101 origin) was straightforward but testing revealed that [[Part:BBa_I50040|BBa_I50040]] was nonfunctional as a replication origin presumably due to introduced mutations.
  
 +
In constructing [[Part:BBa_I50042|BBa_I50042]], the replication origin boundaries were chosen quite liberally (i.e. they might include more sequence than is strictly necessary) in part because [[Part:BBa_I50040|BBa_I50040]] was shown to be non-functional.
  
 +
[[Part:BBa_I50042|BBa_I50042]] is very similar to the replication origin used in Bujard's pZS4Int-1 vector and in Michael Elowitz's repressilator. However, a mutation was introduced to eliminate a SpeI site for compatibility with the BioBrick assembly standard. [[Part:BBa_I50042|BBa_I50042]] is also the origin of replication in pSB4* series plasmids.
  
 
===Source===
 
===Source===
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We constructed [[Part:BBa_I50042|BBa_I50042]] by PCR using [[Part:pSB4A3|pSB4A3]]-[[Part:BBa_P1010|P1010]] as a template and amplification primers I50042-f (<code>5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCTGTCAGACCAAGTTTACGAG-3'</code>) and I50042-r (<code>5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTTACATTGTCGATCTGTTC-3'</code>).
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This part was amplified from pSB4A3-P1010.
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===References===
 
===References===
 +
<biblio>
 +
#Cohen-J-Bacteriol-1977 pmid=334752
 +
#Lutz-Nucleic-Acids-Res-1997 pmid=9092630
 +
#Elowitz-Nature-2000 pmid=10659856
 +
</biblio>

Revision as of 03:39, 25 January 2008

pSC101 origin of replication


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The original design for BBa_I50042 was BBa_I50040. Synthesis of BBa_I50040 (designed pSC101 origin) was straightforward but testing revealed that BBa_I50040 was nonfunctional as a replication origin presumably due to introduced mutations.

In constructing BBa_I50042, the replication origin boundaries were chosen quite liberally (i.e. they might include more sequence than is strictly necessary) in part because BBa_I50040 was shown to be non-functional.

BBa_I50042 is very similar to the replication origin used in Bujard's pZS4Int-1 vector and in Michael Elowitz's repressilator. However, a mutation was introduced to eliminate a SpeI site for compatibility with the BioBrick assembly standard. BBa_I50042 is also the origin of replication in pSB4* series plasmids.

Source

We constructed BBa_I50042 by PCR using pSB4A3-P1010 as a template and amplification primers I50042-f (5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCTGTCAGACCAAGTTTACGAG-3') and I50042-r (5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTTACATTGTCGATCTGTTC-3').

References

<biblio>

  1. Cohen-J-Bacteriol-1977 pmid=334752
  2. Lutz-Nucleic-Acids-Res-1997 pmid=9092630
  3. Elowitz-Nature-2000 pmid=10659856

</biblio>