Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Caleb J. Carr
Date created: 2013-05-07 11:00:00
Date modified: 2015-08-31 04:08:27
Part for inserting modified D-Dogs for Golden Gate Assembly using BsgI
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_J119142_sequence (Version 1) |
Description
This part is a modification of the J100091 construct that removes the C-Dog part and inserts a placeholder fragment with two BsgI sites that both face out from the center of the insert. This part is the precursor to introducing modified C-Dog parts with nested BsgI or BpmI sites to be used in Golden Gate Assembly.This part was designed to complete the library of functional parts being used in the design of a generalized method for optimizing a metabolic pathway.
Notes
To insert this part, oligonucleotides had to be ordered, annealed, and treated with polynucleotide kinase from the BBA prefix to around 110 NT into the mRFP gene where a rogue BsiWI site was discovered. This approach swapped the original ~330 NT from J100091 with a fragment around 220 NT long. However, the mRFP gene was left in tact while the C-Dog element was removed completely.Source
Part is mostly nonsense except for restriction enzyme recognition sequence.| Sequence Annotation | Location | Component / Role(s) |
| Sticky word BsaI (left) J119030 BsaI (right) Sticky word BsgI sticky word (left) "Stuffer" (non-functional) BsgI (left) BsgI (right) BsgI Sticky word (right) mRFP Start Codon Stop Stop | 1,4 6,11 12,45 48,53 55,58 59,60 59,106 75,80 85,90 105,106 106,786 106,108 781,783 784,786 | feature/misc sequence_feature feature/dna sequence_feature feature/promoter promoter feature/dna sequence_feature sequence_feature feature/misc sequence_feature feature/misc feature/dna sequence_feature sequence_feature feature/dna sequence_feature feature/dna feature/misc sequence_feature feature/protein CDS start_codon feature/start stop_codon feature/stop stop_codon feature/stop |