Difference between revisions of "Part:BBa K1985016"
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This part encodes Part:BBa_K1321333 in a pSB1A3 backbone. It was ligated to this backbone as it confers the ampicillin resistance that was needed for in vivo expression for our composite parts: Bba_K1985017, Bba_K1985008, Bba_K1985009, Bba_K1985013, Bba_K1985014, Bba_K1985015. | This part encodes Part:BBa_K1321333 in a pSB1A3 backbone. It was ligated to this backbone as it confers the ampicillin resistance that was needed for in vivo expression for our composite parts: Bba_K1985017, Bba_K1985008, Bba_K1985009, Bba_K1985013, Bba_K1985014, Bba_K1985015. | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K1985016 SequenceAndFeatures</partinfo> | ||
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===Validation=== | ===Validation=== | ||
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[[File:PSB1A3+promoter.jpeg||400px|thumb|centre|Figure 1.1% agarose gel of the restriction digest of BBa_K1985016 in pSB1A3 plasmid backbone with EcoRI and PstI]] | [[File:PSB1A3+promoter.jpeg||400px|thumb|centre|Figure 1.1% agarose gel of the restriction digest of BBa_K1985016 in pSB1A3 plasmid backbone with EcoRI and PstI]] | ||
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Revision as of 22:02, 19 October 2016
AraC-pBAD in pSB1A3
This part encodes Part:BBa_K1321333 in a pSB1A3 backbone. It was ligated to this backbone as it confers the ampicillin resistance that was needed for in vivo expression for our composite parts: Bba_K1985017, Bba_K1985008, Bba_K1985009, Bba_K1985013, Bba_K1985014, Bba_K1985015.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Validation
The plasmid was analysed through a diagnostic double restriction cut, using the enzymes EcoRI and PStI. This was followed by agarose gel electrophoresis. The enzymes cleave the pBAD Ara-C promoter at 1165 bp, with the remainder plasmid being 2114 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct.
