Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Julia Pon
Date created: 2009-10-20 11:00:00
Date modified: 2015-05-08 01:11:08
terminator forward strand B, Biobytes assembly method
| Types | DnaRegion |
| Roles | Other sequence_feature |
| Sequences | BBa_K187090_sequence (Version 1) |
Description
This part can be annealed to the terminator reverse strand (BBa_K187077), to produce a B overhang, which can anneal to the B' end on the 3' end of a part produced from pAB.The biobytes assembly method can be used to rapidly assemble DNA segments in a particular order. The 3' end of constructs made with this method can be annealed to a terminator segment, which can be digested and annealed to the anchor to circularize the construct.
For more details see http://2009.igem.org/Team:Alberta/DNAanchor, or RFC 47
Notes
Once the construct has been completed, i.e. the last Byte has been added, the construct may be released from the beads as is by a simple I-SceI digestion or a USERTM digestion, thus yielding a linear construct. If a circular construct, such as a plasmid, is desired then a final "Terminator" piece must be added. This piece is similar in construction to the Anchor, whereby there is a dsDNA I-SceI recognition sequence with four deoxyuracils incorporated into it on one strand, as well as an A or B 3' overhang. The Terminator binds to the last Byte and release is once again achieved by I-SceI digestion or USERTM digestion. In either case both the Anchor and Terminator develop sticky ends that are complementary to eachother: 4 bases if I-SceI digestion is utilized, or 21 bases if USER is used. USERTM digestion is obviously preferred since 21 bp of interaction will form spontaneously and without ligation, and thus transformation of the construct can proceed immediately.Source
oligonucleotide synthesis| Sequence Annotation | Location | Component / Role(s) |
| should be uracil not thymine should be uracil not thymine should be uracil not thymine should be uracil not thymine | 2,2 8,8 15,15 21,21 | sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc |