Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Malcolm Campbell
Date created: 2011-08-29 11:00:00
Date modified: 2015-08-31 04:08:22
placeholder insert for BsaI Golden Gate Assembly of promoter
| Types | DnaRegion |
| Roles | Other sequence_feature |
| Sequences | BBa_J100028_sequence (Version 1) |
Description
This insert can be placed in any vector that does not contain a BsaI site. BsaI is a type IIs restriction enzyme use in Golden Gate Assembly of multiple inserts. Most versions of ampicillin resistance in plasmids contain a BsaI site. We are putting this insert into a modified pSB1A2 that has its vector BsaI site mutated (see part number ##). This insert can be digested with BsaI and mixed with ligase to make room for a promoter of your choice that has appropriate sticky ends. We are using this insert in combination with new promoters assembled from oligonucleotides (oligos). When assembled, the new promoter would have a four base overhang of 5' GCTG 3' on the left side of the top strand and a four base overhang of 3' CGCC 5' on the right side of the bottom strand:5' CGACNNN...NNN 3'
3' NNN...NNNCGCC 5'
This insert contains BBa biobrick prefix, BbsI site, 4 base overhang followed by BsaI site cutting to the left, transcriptional terminator B0014, BsaI site that cuts to the right, 4 base overhang, BbsI site, RBS B0034, mRFP part number ##) and ending in BBa biobrick suffix.
Notes
Had to avoid BsaI sites and BbsI sites as well as all biobrick restriction sites.Source
Synthesized de novo using DNA sequences from parts in the registry. Typical scars are absent from this DNA.| Sequence Annotation | Location | Component / Role(s) |
| prefix suffix B0034 BbsI cuts left start double stop BsaI cuts left misc BsaI cuts right misc BbsI cuts right mRFP E1010 | 1,21 858,877 158,169 23,28 176,178 851,856 34,39 29,32 135,140 142,145 146,151 176,856 | feature/BioBrick engineered_region engineered_region feature/BioBrick feature/rbs ribosome_entry_site feature/binding non_covalent_binding_site start_codon feature/start stop_codon feature/stop non_covalent_binding_site feature/binding sequence_feature feature/misc feature/binding non_covalent_binding_site feature/misc sequence_feature non_covalent_binding_site feature/binding feature/cds CDS |