Device

Part:BBa_J119394

Designed by: Athena J. Davis   Group: Eckdahl Lab   (2016-06-23)
Revision as of 13:59, 28 June 2016 by Eckdahl (Talk | contribs)


tClone TetA+SacB: Device for Testing Transciptional Terminators and Riboswitches via GGA

tClone TetA+SacB can be used to clone and test new transcriptional terminators and riboswitches that function by antitermination without gel purification or other preparation of DNA. A new terminator or riboswitch can be derived from synthetic oligos, PCR, or a plasmid clone and cloned into tClone TetA with Golden Gate Assembly (GGA) using BsaI and ligase. For proper assembly, see BBa_J119367. TetA encodes tetracycline resistance, and expression of SacB causes cell death in the presence of sucrose. Whether or not transcription proceeds to the TetA gene is determined by the strength of the terminator or whether termination or antitermination occurs at the riboswitch.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1095
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1241
    Illegal BamHI site found at 3438
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1267
    Illegal NgoMIV site found at 1635
    Illegal NgoMIV site found at 1795
    Illegal AgeI site found at 3385
    Illegal AgeI site found at 3412
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 849
    Illegal BsaI.rc site found at 42


[edit]
Categories
Parameters
None