Difference between revisions of "Part:BBa K341456:Design"

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===References===
 
===References===
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Thomas E. Kuhlman and Edward C. Cox.(2009)Site-specific chromosomal integration of large synthetic constructs.Nucleic Acids Research, 2009, 1–10

Latest revision as of 02:52, 28 October 2010

Kanamycin resistance(with Promoter) + I-sceI site+ Recombination site


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 893
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 229
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 169
    Illegal SapI site found at 379
    Illegal SapI site found at 1225


Design Notes

This part involves DraIII cutting site at two ends of the unit, this cutting site is compatible with BBF RFC 61

Thsi part is uesd as a Insertion Fragment of Donor Plasmid in the In-vivo Recombination System of iGEM Tsinghua 2010 Project.

Donor Plasmid contains several Insertion Fragments and each Insertion Fragment contains following five parts:

1) one 15bp-long restriction enzyme I-scel recognition site

2) one 25bp-long random sequence

3) one fragment for insertion and recombination

4) one 25bp-long random sequence

5) one 15bp-long restriction enzyme I-scel recognition site(in correspondence with 1)

Donor plasmid contains several Insertion Fragments. Based on the flanking landing pad sequence, we can ascribe Insertion Fragments to the same group as long as the flanking landing pad sequences of those Insertion Fragments are the same.

In order to achieve different goals, we design different Donor Plasmids, different Donor Plasmids contain different number of Insertion Fragments, which belong to different groups.

Source

Kanamycin resistance gene is form plasmid "pKD13" which is generously provided by Guoqiang CHEN of Microorganism Lab in Tsinghua University

References

Thomas E. Kuhlman and Edward C. Cox.(2009)Site-specific chromosomal integration of large synthetic constructs.Nucleic Acids Research, 2009, 1–10