Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Chris French, Allan Crossman, Sylvia Ispasanie
Date created: 2011-06-27 11:00:00
Date modified: 2015-05-08 01:12:33
Plac + lacZ
with BglII site
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_K523000_sequence (Version 1) |
Description
This is based on part BBa_J33207. Like that part, it encodes the lac promoter and the N-terminal 76 amino acid residues of LacZ, which produce a peptide that complements the lacZ-delta-M15 mutation in common lab strains of E. coli. If grown on Xgal and IPTG, colonies with this part will be blue.The first four bases of the sequence add a BglII site (BglII sites have 6 bases, but the first 2 are provided by the standard BioBrick prefix). This enables an easy method of using PCR to create new BioBricks (e.g. from natural sources): the forward primer for the new BioBrick should have a BglII site added, and the reverse primer should have a SpeI site added.
The PCR product and the plasmid containing this part can then both be digested with BglII and SpeI. After mixing and ligation, plasmids that have successfully gained the new BioBrick will be white, while those still containing the lacZ part will be blue (on Xgal+IPTG plates).
One might ask: why not use the standard flanking BioBrick sites XbaI and SpeI for this procedure? The answer is that these produce compatible sticky ends and so, upon digestion, the new part will circularise.
Notes
Note that the BglII (agatct) site overlaps the BioBrick prefix. Since the prefix ends "ag", only "atct" is included in the BioBrick proper.Source
Based on part BBa_J33207, which was amplified from E. coli BL21 genomic DNA using primers based on published sequence (Genbank accession J01636, gi:146575).| Sequence Annotation | Location | Component / Role(s) |
| BglII rbs LacZ' CAP binding site -35 -10 LacI binding site ATG TGA | 1,4 315,318 326,556 204,241 253,258 276,281 288,308 326,326 554,554 | sequence_feature feature/misc ribosome_entry_site feature/rbs CDS feature/cds sequence_feature feature/misc promoter feature/promoter feature/promoter promoter sequence_feature feature/misc start_codon feature/start feature/stop stop_codon |