Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Richard Partridge-Hicks and Maria Kowal
Date created: 2010-10-06 11:00:00
Date modified: 2015-05-08 01:11:59
lac promoter + luxAB
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K322140_sequence (Version 1) |
Description
The first part (submitted in pSB1A2) consists of the lac promoter and lacZ' gene, encoding the N-terminal 76 amino acid residues of LacZ, sufficient to complement the lacZ-delta-M15 mutation for blue-white selection on Xgal plates. A SacI site has been introduced at the 3' end, overlapping the XbaI site of the Biobrick prefix. This is designed to be used as a cloning vector for making new biobricks. PCR primers can be designed with a SacI site in one primer and an SpeI site in the other. This removes the necessity for an excessively long non-complementary tail on one primer bearing either the full biobrick prefix or suffix. The PCR product can then be digested with SacI and SpeI for insertion into this plasmid, replacing the Plac-lacZ' cassette. Recombinant plasmids will then be white on IPTG/Xgal plates, whereas any that still contain the original insert will be blue. We have used this strategy to prepare several biobricks, including BBa_J33204, which contains the xylE gene encoding catechol-2,3-dioxygenase.For the second part, which is LuxAB coding, has a known gene sequence of luxCDAB(F)E, where lux A and lux B code for the components of luciferase, and the lux CDE codes for a fatty acid reductase complex that makes the fatty acids necessary for the luciferase mechanism (Meighen, 1991).
Notes
J33207 Note that the SacI site overlaps the SpeI site. The Biobrick prefix ends ...TCTAGAG. When this is added to the CTC at the start of the sequence shown here, the SacI site, GAGCTC, is generated.K322139 Notes: we prepared this for two reasons: firstly to resolve an issue with the SpeI site in our submitted version of K216008 (for some reason, every clone we sequenced seemed to have an error, not always the same error, in the suffix SpeI site), and to put it in pSB1C3, since this seems to be the new standard vector for the Registry.
Source
J33207 The DNA was amplified from E. coli BL21 genomic DNA using primers based on published sequence (Genbank accession J01636, gi:146575). The annotation shown here is based on that associated with this Genbank entry. The sequence shown here is derived by sequencing the construct.References
K322139 Source: genomic DNA of Xenorhabdus (Photorhabdus) luminescens Hb (actually, we used our BioBrick from last year, BBa_K216016 as template).
| Access | Instance | Definition |
| public public | BBa_J33207 BBa_K322139 | BBa_J33207 BBa_K322139 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J33207 BBa_K322139 | 1,600 609,2718 | BBa_J33207 BBa_K322139 |