Reporter

Part:BBa_K341002:Design

Designed by: Sheng WANG   Group: iGEM10_Tsinghua   (2010-10-20)

promoter+tet-SDS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1147
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1-25： LP1

26-43： I-scel

50-80：Plac

115-1320： Tet

1321-1338：I-Scel

1339-1363： LP2

This part is used as a "Landing Pad"in the In-vivo Recombination System. This ”Landing Pad(LP)” should be transformed into the specific sites of E coli via Att recombination.

Landing pad sequence consists of the following parts(from 5’ to 3’ in the sequence):

1）25bp-long random sequence

2）15bp-long recognition sequence of restriction enzyme I-scel

3) antibiotic resistance gene used for antibody selection

4) 15bp-long recognition sequence of restriction enzyme I-scel(corresponding to 2)

5) 25bp-long random sequence (corresponding to 1)

After integrating DNA segment of Landing pad into the genome of E coli, we completed the genetic engineering of the genome of E coli. All the five parts of landing pad are designed for subsequent recombination. Besides, in order to achieve different recombination goals, we designed several landing pads of different sequences.

Source

These sequence is from Vecter ptks-cs which is generously provided by Thomas E. Kuhlman and Edward C. Cox of the Department of Molecular Biology, Princeton University.

References

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