Difference between revisions of "Part:BBa J119394"

Latest revision as of 19:39, 28 June 2016

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 and SacB genes 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

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 <partinfo>BBa_J119394 short</partinfo>
-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.
+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 and SacB genes is determined by the strength of the terminator or whether termination or antitermination occurs at the riboswitch.
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