DNA

Part:BBa_K617003:Design

Designed by: Will Jones   Group: iGEM11_UIUC-Illinois   (2011-09-28)

R6K 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

An R6K conditional origin of replication is located between the biobrick prefix and suffix site of the pSB1C3 backbone. The R6K origin is from the CRIM pAH125 plasmid. This origin is only functional when the trans-acting protein product of the pir gene is expressed (Metcalf et al., 1994). This part may be used to create new vector backbones.

The Sequence of R6K has been confirmed through sequencing by ACGT Inc. using the biobrick VF2 and VR primers.

Source

K617003 was pcred off the CRIM pAH125 plasmid, material which was designed by Haldimann and Wanner (2001) at Purdue University, and was subsequently cloned via 3A assembly into a pSB1C3 vector supplied by the Registry of Standard Biological Parts.

References

1. Haldimann, A., Wanner, B. L. 2001. Conditional-Replication, Integration, Excision, and Retrieval Plasmid-Host Systems for Gene Structure-Function Studies of Bacteria. Journal of Bacteriology. 183:21 6384-6393.

2. Metcalf, W. W., Weihong, J., Wanner, B. L. 1994. Use of the rep technique for allele replacement to construct new Escherichia coli hosts for maintenance of R6Kϒ origin plasmids at different copy numbers. Gene. 138: 1-7.