Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Yoshio Takata
Date created: 2016-10-05 11:00:00
Date modified: 2016-10-11 02:10:58
cold inducible promoter (Pcold)
| Types | DnaRegion |
| Roles | promoter Regulatory |
| Sequences | BBa_K1949000_sequence (Version 1) |
Description
This new promoter, a cold inducible promoter (we call this Pcold) consists of the cspA promoter, Cold Box, 5???-UTR, RBS and DB. This promoter is used to effectively produce proteins at low temperatures.Biobrick Tips
This part is not able to be used for most common assembly, because restriction enzyme digestion with XbaI and SpeI generates an unexpected stop codon. Therefore, this part do not meet the criteria of basic parts construction. Our team generated a unique digestion site, BamHI at the upstream of the suffix. We recommend to use this BamHI site for cloning.
Characterization
Notes
We applied a point mutation to the Pcold in order to be easy to clone it. We changed the 178th base C to A. An article which we referred describes as stated below.???we found occasionally that C???A base substitution at nucleotide number 532 in the 5???-UTR region (nucleotide number is according to GenBank Accession No. M30139) diminished this growth inhibition effect (data not shown), allowing easy cloning of the genes of in- terest into MCS. We do not know why this mutation diminished growth inhibition effect but the mutation did not affect productivity of proteins (data not shown).???
(Nakashima,N and Tamura,T. 2004. Cell-free protein synthesis using cell extract of Pseudomonas fluorescens and CspA promoter. Biochemical and Biophysical Research Communications 319 (2004) 672)
Source
We artificial synthesized this.| Sequence Annotation | Location | Component / Role(s) |
| mutation A to C BamHI | 177,177 308,313 | sequence_alteration feature/mutation feature/misc sequence_feature |