BBa_K524201 1 BBa_K524201 Modified ccdB for plasmid assembly on standard vectors 2012-08-18T11:00:00Z 2015-06-15T12:38:08Z Modified P1010 was constructed from BBa_P1010 (ccdB cell death gene) on pSB1C3 by PCR amplification. This part was modified from the existing BioBrick BBa_P1010 (ccdB cell death gene), flanked by restriction sites AvrII (followed after RFC10 prefix) and NheI (preceding RFC10 suffix). One of the many ways to construct plasmids from BioBricks would be to perform standard assembly on a standard vector, excise the insert out using XbaI and SpeI, and then allow the linearized plasmid to self ligate. Such strategy usually involves the purification of desired fragment by electrophoresis followed by gel purification. If the insert is larger than the standard vector, the excised gel fragment might be contaminated by the vector. Contaminant standard vectors would self-ligate and possess higher transformation efficiency than the desired constructed plasmid. In cases where such 'co-transformation' are not desired, the modified ccdB cell death gene can be added at either end of insert by standard assembly (in such situation a ccdB tolerant strain should be used to harbor the construct), and the intended fragment can be excised using either a combination of XbaI + AvrII or SpeI + NheI. As transformation proceeds, any undesired fragments carrying the ccdB part would kill the host cells and the remaining population would only hold the plasmid of interest. true false _689_ 4206 8758 9 Not in stock false The AvrII site and NheI site were encouraged to exclude from standard biobricks. The addition of two sites but not one is the result considering that either site might be unavoidable in some parts. The arrangement that the AvrII site precedes the NheI site is plainly arbitrary. false HO Yuan Heng Trevor annotation2179951 1 AvrII site range2179951 1 1 6 annotation2179954 1 ccdA range2179954 1 342 562 annotation2179955 1 BBa_P1010 range2179955 1 7 681 annotation2179952 1 NheI site range2179952 1 682 687 annotation2179973 1 -10 range2179973 1 610 615 annotation2180019 1 -35 range2180019 1 624 629 annotation2179953 1 ccdB range2179953 1 35 340 BBa_K524201_sequence 1 cctaggactggctgtgtataagggagcctgacatttatattccccagaacatcaggttaatggcgtttttgatgtcattttcgcggtggctgagatcagccacttcttccccgataacggagaccggcacactggccatatcggtggtcatcatgcgccagctttcatccccgatatgcaccaccgggtaaagttcacgggagactttatctgacagcagacgtgcactggccagggggatcaccatccgtcgcccgggcgtgtcaataatatcactctgtacatccacaaacagacgataacggctctctcttttataggtgtaaaccttaaactgcatttcaccagcccctgttctcgtcagcaaaagagccgttcatttcaataaaccgggcgacctcagccatcccttcctgattttccgctttccagcgttcggcacgcagacgacgggcttcattctgcatggttgtgcttaccagaccggagatattgacatcatatatgccttgagcaactgatagctgtcgctgtcaactgtcactgtaatacgctgcttcatagcatacctctttttgacatacttcgggtatacatatcagtatatattcttataccgcaaaaatcagcgcgcaaatacgcatactgttatctggcttttagtaagccggatccacgcgtgctagc igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z