Device

Part:BBa_K3011007:Design

Designed by: Pinaki Chanda   Group: iGEM19_IISER_Kolkata   (2019-10-15)
Revision as of 08:10, 20 October 2019 by The genetic bot (Talk | contribs)


Nitric Oxide sensor

Constitutive Promoter (BBa_J23100) – RBS (BBa_B0032) – NsrR (BBa_K1682011) – Terminator (BBa_B0010) – PyeaR promoter (BBa_K216005) – RBS (BBa_B0032) – mRFP (BBa_E1010) – Terminator (BBa_B0012)

NsrR (BBa_K1682011) is a Nitric Oxide (NO) sensitive transcription repressor. This construct produces NsrR constitutively; in absence of NO, NsrR binds to the NsrR binding site present on the PyeaR promoter (BBa_K216005) and repress the transcription of mRFP (BBa_E1010). In presence of NO, NsrR binds to NO due to which NsrR can’t bind to the DNA (thus can’t repress gene expression) and mRFP will produce.

NsrR contains a Helix Turn Helix (HTH) motif and Fe-S cluster ([2Fe-2S] or [4Fe-4S]). In absence of NO, NsrR binds to a particular DNA sequence by using HTH motif (as a homodimer) and repress gene expression. In presence of NO, Fe-S cluster reacts with NO to form dinitrosyl iron complex, due to the formation of this complex NsrR can’t bind to that particular sequence anymore. Depending upon the sequence of NsrR binding site, the number of NO molecules (per cluster) which is required for abolishing DNA binding vary (2-10 NO / cluster)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 202
    Illegal AgeI site found at 1222
    Illegal AgeI site found at 1334
  • 1000
    COMPATIBLE WITH RFC[1000]