Difference between revisions of "Part:BBa K4364000"
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This part can be used to create a T-vector that is compatible with various TA cloning protocols. In order to do this, it should be cloned in a suitable pSB vector, this plasmid must be extracted and then treated with the restriction endonuclease AhdI. It will produce a linearized vector with single T overhangs. Moreover, the TA-cassette is integrated into the reading frame of mRFP1 under the control of the lac promoter. The empty vector produces notable red-colored colonies. If the TA cloning is successful the positive transformants will be white. | This part can be used to create a T-vector that is compatible with various TA cloning protocols. In order to do this, it should be cloned in a suitable pSB vector, this plasmid must be extracted and then treated with the restriction endonuclease AhdI. It will produce a linearized vector with single T overhangs. Moreover, the TA-cassette is integrated into the reading frame of mRFP1 under the control of the lac promoter. The empty vector produces notable red-colored colonies. If the TA cloning is successful the positive transformants will be white. | ||
Revision as of 08:22, 12 October 2022
mRFP1-based cassette for TA cloning with dual T7 promotors
This part can be used to create a T-vector that is compatible with various TA cloning protocols. In order to do this, it should be cloned in a suitable pSB vector, this plasmid must be extracted and then treated with the restriction endonuclease AhdI. It will produce a linearized vector with single T overhangs. Moreover, the TA-cassette is integrated into the reading frame of mRFP1 under the control of the lac promoter. The empty vector produces notable red-colored colonies. If the TA cloning is successful the positive transformants will be white.
Please be careful if you transfer this part to pSB_A_ plasmid backbones - the beta-lactamase encoded by these vectors contains an AhdI site that should be destroyed first.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 199
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 207
Illegal BamHI site found at 316 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 906
Illegal AgeI site found at 1018 - 1000COMPATIBLE WITH RFC[1000]