Difference between revisions of "Part:BBa K510048:Design"
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===References=== | ===References=== | ||
Ella Gringauz, Karina A. Orle, Candace S. Waddell, and Nancy L. Craig (1988). Recognition of Escherichia coli attTn7 by Transposon Tn7: Lack of Specific Sequence Requirements at the Point of Tn7 Insertion. Journal of Bacteriology, June 1988, p. 2832-2840. | Ella Gringauz, Karina A. Orle, Candace S. Waddell, and Nancy L. Craig (1988). Recognition of Escherichia coli attTn7 by Transposon Tn7: Lack of Specific Sequence Requirements at the Point of Tn7 Insertion. Journal of Bacteriology, June 1988, p. 2832-2840. | ||
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Candace S. Waddellt and Nancy L. Craig (1989). Tn7 transposition: Recognition of the attTn7 target sequence. Proc. Natl. Acad. Sci. USA Vol. 86, pp. 3958-3%2, June 1989, Biochemistry. Joseph E. Peters and Nancy L. Craig (2001). Tn7: smarter than we thought. Nature reviews, November 2001, Vol. 2, 806. | Candace S. Waddellt and Nancy L. Craig (1989). Tn7 transposition: Recognition of the attTn7 target sequence. Proc. Natl. Acad. Sci. USA Vol. 86, pp. 3958-3%2, June 1989, Biochemistry. Joseph E. Peters and Nancy L. Craig (2001). Tn7: smarter than we thought. Nature reviews, November 2001, Vol. 2, 806. | ||
Latest revision as of 16:01, 22 October 2011
pUC18R6KT-miniTn7BB-Gm-attTn7
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4528
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4528
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4534 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4528
Illegal BglII site found at 3212
Illegal BglII site found at 3483
Illegal BglII site found at 3769 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4528
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 4528
Illegal XbaI site found at 4543
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1854
Illegal SapI site found at 518
Design Notes
The portable attTn7 (BBa_K510022) BioBrick was inserted within the BCS of pUC18R6KT-miniTn7BB-Gm (BBa_K510012) by EcoRI and PstI clonning.
Source
The pUC18R6KT vector was amplified by PCR using as template a pTNS2 plasmid (GenBank accession number: AY884833) provided by Herbert P. Schweizer. The primers were designed to flanked the pUC18R6KT with SfiI restriction sites and allow the insertion of the mini-Tn7-Gm. Primers: attcGGCCTAGGCGGCCgtcgttttacaacgtcgtgac and attcGGCCGCCTAGGCCggaagcataaagtgtaaagcctg.
The miniTn7BB-Gm minitransposon was synthesized commercially, and then digested at the flanking SfiI sites and cloned into SfiI-digested pUC18R6KT PCR products.
The attTn7 was obtained from the purified genome of E. coli MC4100 by PCR (using a high accuracy Pfu polymerase) with the follow primers: gaattcgcggccgcttctagagTGCAGCTGCTGGCTTACCATG and ctgcagcggccgctactagtaACATGGAGTTGGCAGGATGTTTG.
References
Ella Gringauz, Karina A. Orle, Candace S. Waddell, and Nancy L. Craig (1988). Recognition of Escherichia coli attTn7 by Transposon Tn7: Lack of Specific Sequence Requirements at the Point of Tn7 Insertion. Journal of Bacteriology, June 1988, p. 2832-2840.
Candace S. Waddellt and Nancy L. Craig (1989). Tn7 transposition: Recognition of the attTn7 target sequence. Proc. Natl. Acad. Sci. USA Vol. 86, pp. 3958-3%2, June 1989, Biochemistry. Joseph E. Peters and Nancy L. Craig (2001). Tn7: smarter than we thought. Nature reviews, November 2001, Vol. 2, 806.