Difference between revisions of "Part:BBa J119374"

Latest revision as of 17:53, 13 November 2014

tClone TetA: Device for Testing Transciptional Terminators and Riboswitches via Golden Gate Assembly

tClone TetA 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. 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
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
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 849
Illegal BsaI.rc site found at 42

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 __NOTOC__
 <partinfo>BBa_J119374 short</partinfo>
-tClone is a construction intermediate that can be used to build devices that will allow users to clone and test new transcriptional terminators and riboswitches that function by antitermination without gel purification or other preparation of DNA. Use of BioBrick cloning to add the TetA gene into the suffix of tClone produce tClone TetA, which can be used to measure new transcriptional terminators and riboswitches. tClone vectors can be used as destination vectors for Golden Gate Assembly (GGA) using BsaI and ligase. A new terminator or riboswitch can be derived from synthetic oligos, PCR, or a plasmid clone. For proper assembly, see BBa_J119367. 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. The part incorporates the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.
+tClone TetA 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. 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.
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