Composite

Part:BBa_K1937004:Design

Designed by: Oumnia KARIM   Group: iGEM16_Toulouse_France   (2016-10-13)
Revision as of 17:24, 13 October 2016 by Oumniak (Talk | contribs)


pNagA-RFP in pSBBs0K-mini plasmid


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 1257
    Illegal suffix found in sequence at 1285
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1257
    Illegal NheI site found at 201
    Illegal NheI site found at 1279
    Illegal SpeI site found at 1286
    Illegal PstI site found at 1300
    Illegal NotI site found at 1263
    Illegal NotI site found at 1293
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1257
    Illegal BglII site found at 999
    Illegal BglII site found at 5814
    Illegal BamHI site found at 1330
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 1257
    Illegal suffix found in sequence at 1286
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 1257
    Illegal XbaI site found at 1272
    Illegal SpeI site found at 1286
    Illegal PstI site found at 1300
    Illegal NgoMIV site found at 2877
    Illegal AgeI site found at 761
    Illegal AgeI site found at 873
    Illegal AgeI site found at 5425
    Illegal AgeI site found at 6387
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1055
    Illegal BsaI.rc site found at 2701
    Illegal BsaI.rc site found at 4140
    Illegal BsaI.rc site found at 6656
    Illegal SapI site found at 1618
    Illegal SapI.rc site found at 5638


Design Notes

The part includes the pNagA promoter and its RBS (position 3,595,156 to 3,595,356 of the Bacillus genome). The promoter controls the expression of the RFP fluorescent reporter gene. A NheI restriction site has been placed before the RFP ORF to facilitate promoter swapping.This part is a subcloning of BBa_K1937003 pNagA part in pSBBS0K_mini plasmid (BBa_K1937001). Here is a map of the gene:

BBa K1937004-map.png


Source

The part include the pNagA promoter and its RBS (position 3,595,156 to 3,595,356 of the Bacillus genome). The promoter controls the expression of the RFP fluorescent reporter gene. A NheI restriction site has been placed before the RFP ORF to facilitate promoter swapping.This part is a subcloning of BBa_K1937003 pNagA part in pSBBS0K_mini plasmid (BBa_K1937001).

In bacillus subtilis, this promoter has been reported to induce expression in presence of N-acetylglucosamine 6-phosphate.


References