Transposable piece of a transposon bearing suicide plasmid, containing chloramphenicol resistance, an origin of replication for decoding, and a selectable gentimicin marker that only activates when the transposon lands in a transcribed region.
Construction intermediate for a transposon selectable for insertion into a transcribed region.
Intermediate for a transposon with a chloramphenicol resistance marker and a replication origin inside of the transposed region.
Overlap extension of synthetic oligonucleotides
DNA substrate for Tn5 transposase. Flanking a sequence with J61021 on the 5' end and J61022 on the 3' end makes it a functional transposon.
DNA substrate for Tn5 transposase. Flanking a sequence with J61021 on the 5' end and J61022 on the 3' end makes it a functional transposon. pSB1A2-Bca1013
Gentamicin resistance gene including a medium/weak ribosome binding site, but NO promoter. Intended for use in transposons, to select for elements that fall inside of a transcribed region. The RBS does not appear to be strong enough for this purpose.
The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.
PCR ca1016F/R on pKD3 (934 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Products are Bca1015 and Bca1016 ---- ca1016F Forward EcoRI to biobrick CAT of pKD3 gacttgaattcgcggccgcttctagaggaataggaacttcatttaaatg ca1016R Reverse SpeI to biobrick CAT of pKD3 ccgctactagtggcgcgcctacctgtgacgg
Chloramphenicol resistance gene including its native promoter and ribosome binding site. Confers resistance to 25 ug/mL chloramphenicol. {pSB1A2-Bca1016}
PCR ca1011F/R on pG80ko (445 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Product is Bca1011 ---- ca1011F Forward Biobricking of R6K GGACTgaattcgcggccgcttctagagtgattcgcacgggcccatg ca1011R Reverse biobricking of R6K ccgctactagtaGCAGTTCAACCTGTTGATAG
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}
Extend ca1010F/R (73 bp, EcoRI/SpeI) Sub into pSB1A2-I13522 (EcoRI/SpeI) Product is pSB1A2-Bca1010 ---- ca1010F Forward (universal biobrick) EcoRI for FRT GGACTgaattcgcggccgcttctagag ca1010R Reverse SpeI oligo for FRT cctatactagtagaagttcctattctctaAaaagtataggaacttcctctagaagcggccgcg
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.
Conversion of the iGEM parts registry to SBOL2.1