Cold shock

AmpR promoter

The AmpR promoter is a crucial regulatory element located in the upstream of the AmpR gene that controls the expression of the AmpR gene, responsible for initiating beta-lactamase expression.

AmpR

The AmpR gene encodes the beta-lactamase enzyme. It helps the bacteria provide resistance to ampicillin.

Ori (origin of replication)

The initiation of plasmid DNA replication.

cspA promoter, 5’-UTR, and 3’-UTR

The cspA promoter is activated in low temperature conditions in E. coli.

TEE (translation enhancing element)

TEE is responsible for enhancing the translation efficiency of mRNA resulting in an increase in protein expression.

otsA

OtsA, trehalose-6-phosphate synthase (TPS), is an enzyme involved in the trehalose synthesis pathway. It catalyzes the synthesis of trehalose-6-phosphate from UDP-glucose and glucose-6-phosphate.

FLAG tag

A protein tag (DYKDDDDK) fused in the C-terminus of OtsA.

T2A

T2A is a self-cleavage peptide that allows the co-expression of multiple genes from a single mRNA transcription.

otsB

OtsB, trehalose 6-phosphate phosphatase (TPP), is an enzyme involved in the trehalose synthesis pathway. It catalyzes the synthesis of trehalose from trehalose-6-phosphate.

Myc tag

A protein tag (EQKLISEEDL) fused in the C-terminus of OtsB.

EGFP (enhanced green fluorescent protein)

EGFP is an enhanced version of the green fluorescent proteins that exhibits green fluorescence when expressed. It has been widely used in the field of synthetic biology as a reporter gene to observe gene expression.