BBa_J61027 1 BBa_J61027 [FRT][R6K][CmR][FRT] 2006-09-20T11:00:00Z 2015-08-31T02:02:59Z standard biobrick assembly Cassette for knockins or knockouts with chloramphenicol selection and a silent origin of replication. The cassettes can be removed by introduction of flp recombinase expressing helper plasmid pCP20 leaving behind a single FRT site scar. Digestion of the genomic DNA containing these cassettes followed by ligation and transformation into a pir116 or pir+ strain allows capture of the cassette as a high or medium-copy plasmid. pSB1A2-Bca9008 false false _95_ 0 483 95 It's complicated false N/A false John Anderson component1902244 1 BBa_J61001 component1902246 1 BBa_J61000 component1902242 1 BBa_J61020 component1902247 1 BBa_J61020 annotation1902247 1 BBa_J61020 range1902247 1 1359 1392 annotation1902242 1 BBa_J61020 range1902242 1 1 34 annotation1902246 1 BBa_J61000 range1902246 1 457 1350 annotation1902244 1 BBa_J61001 range1902244 1 43 448 BBa_J61001 1 R6K [R6K] Origin of replication 2006-07-30T11:00:00Z 2015-08-31T02:02:59Z <tt> PCR ca1011F/R on pG80ko (445 bp, EcoRI/SpeI)<br> Sub into pSB1A2-I13522 (EcoRI/SpeI)<br> Product is Bca1011<br> ---- ca1011F Forward Biobricking of R6K<br> GGACTgaattcgcggccgcttctagagtgattcgcacgggcccatg<br> ca1011R Reverse biobricking of R6K<br> ccgctactagtaGCAGTTCAACCTGTTGATAG<br> </tt> Released HQ 2013 R6K origin of replication requires a pir+ or pir116 strain for replication in the absence of a second origin. In most E. coli strains, this is a silent feature. {pSB1A2-Bca1011} false false _95_ 0 483 95 In stock false N/A true John Anderson annotation1892271 1 R6K range1892271 1 27 406 BBa_J61000 1 CmR chloramphenicol resistance cassette 2006-07-30T11:00:00Z 2015-08-31T02:02:59Z <tt> PCR ca1016F/R on pKD3 (934 bp, EcoRI/SpeI)<br> Sub into pSB1A2-I13522 (EcoRI/SpeI)<br> Products are Bca1015 and Bca1016<br> ---- ca1016F Forward EcoRI to biobrick CAT of pKD3<br> gacttgaattcgcggccgcttctagaggaataggaacttcatttaaatg<br> ca1016R Reverse SpeI to biobrick CAT of pKD3<br> ccgctactagtggcgcgcctacctgtgacgg<br> </tt> Chloramphenicol resistance gene including its native promoter and ribosome binding site. Confers resistance to 25 ug/mL chloramphenicol. {pSB1A2-Bca1016} false false _95_ 0 483 95 It's complicated false N/A true John Anderson annotation1892270 1 CmR range1892270 1 30 687 BBa_J61020 1 FRT [FRT] 2006-09-20T11:00:00Z 2015-08-31T02:02:59Z <tt> Extend ca1010F/R (73 bp, EcoRI/SpeI)<br> Sub into pSB1A2-I13522 (EcoRI/SpeI)<br> Product is pSB1A2-Bca1010<br> ---- ca1010F Forward (universal biobrick) EcoRI for FRT<br> GGACTgaattcgcggccgcttctagag<br> ca1010R Reverse SpeI oligo for FRT<br> cctatactagtagaagttcctattctctaAaaagtataggaacttcctctagaagcggccgcg<br> </tt> Site for recombination by flp recombinase. Genes flanked by FRT sites (oriented in the same direction) in the genome can be excised with the introduction of flp recombinase-expressing helper plasmid pCP20. Note that the original FRT sequence contained an XbaI site that has been removed with a point mutation for compatibility with standard assembly. See Datsenko and Wanner for details of its use in markerless knockouts and knockins. false false _95_ 0 483 95 It's complicated false N/A false John Anderson BBa_J61027_sequence 1 gaagttcctatactttttagagaataggaacttctactagagtgattcgcacgggcccatggctaattcccatgtcagccgttaagtgttcctgtgtcactcaaaattgctttgagaggctctaagggcttctcagtgcgttacatccctggcttgttgtccacaaccgttaaaccttaaaagctttaaaagccttatatattcttttttttcttataaaacttaaaaccttagaggctatttaagttgctgatttatattaattttattgttcaaacatgagagcttagtacgtgaaacatgagagcttagtacgttagccatgagagcttagtacgttagccatgagggtttagttcgttaaacatgagagcttagtacgttaaacatgagagcttagtacgtgaaacatgagagcttagtacgtactatcaacaggttgaactgctactagaggaataggaacttcatttaaatggcgcgccttacgccccgccctgccactcatcgcagtactgttgtattcattaagcatctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacgtaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgacaactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgccgatcaacgtctcattttcgccaaaagttggcccagggcttcccggtatcaacagggacaccaggatttatttattctgcgaagtgatcttccgtcacaggtaggcgcgctactagaggaagttcctatactttttagagaataggaacttc BBa_J61001_sequence 1 tgattcgcacgggcccatggctaattcccatgtcagccgttaagtgttcctgtgtcactcaaaattgctttgagaggctctaagggcttctcagtgcgttacatccctggcttgttgtccacaaccgttaaaccttaaaagctttaaaagccttatatattcttttttttcttataaaacttaaaaccttagaggctatttaagttgctgatttatattaattttattgttcaaacatgagagcttagtacgtgaaacatgagagcttagtacgttagccatgagagcttagtacgttagccatgagggtttagttcgttaaacatgagagcttagtacgttaaacatgagagcttagtacgtgaaacatgagagcttagtacgtactatcaacaggttgaactgc BBa_J61000_sequence 1 gaataggaacttcatttaaatggcgcgccttacgccccgccctgccactcatcgcagtactgttgtattcattaagcatctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacgtaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgacaactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgccgatcaacgtctcattttcgccaaaagttggcccagggcttcccggtatcaacagggacaccaggatttatttattctgcgaagtgatcttccgtcacaggtaggcgcgc BBa_J61020_sequence 1 gaagttcctatactttttagagaataggaacttc 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