Reporter

Part:BBa_K1151011

Designed by: Gian Piero Lazzari   Group: iGEM13_UniSalento_Lecce   (2013-09-23)

GFP nickel regulated 3

This is a nickel sensitive system driven by the nikR-exbB intergenic region (BBa_K1151002) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR (BBa_K1151000) as trascription factor.

Boiling prep and digestion with restriction enzymes

Once inserted the construct in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check.

                         EPEP3Unisalento.jpg

Figure 1: Digestion result of boiling prep plasmids.

PCR with VF2 and VR2 primers

                PCR77Unisalento.jpg

Figure 2: PCR results.


Sequence and Features


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 919


[edit]
Categories
Parameters
n/aGFP nickel regulated 3