User guide

Map and features of pcDNA

™

3.3-TOPO

vector, continued

Features

The pcDNA

™

3.3-TOPO

vector contains the following elements. Features have

been functionally tested, and the vectors have been fully sequenced.

Feature Benefit

Full length human

cytomegalovirus (CMV)

immediate-early

promoter/enhancer

Allows efficient, high-level expression of

your recombinant protein (Andersson et al.,

1989; Boshart et al., 1985; Hennighausen &

Fleckenstein, 1986; Nelson et al., 1987).

CMV forward primer Allows sequencing of the insert.

TOPO

Cloning site Allows insertion of your PCR product.

TK polyA reverse primer Allows sequencing of the insert.

Herpes Simplex Virus

Thymidine Kinase (TK)

polyadenylation signal

Allows efficient transcription termination

and polyadenylation of mRNA (Cole &

Stacy, 1985).

SV40 early promoter and origin Allows efficient, high-level expression of the

neomycin resistance gene and episomal

replication in cells expressing the SV40 large

T antigen.

Neomycin resistance gene Allows selection of stable transfectants in

mammalian cells (Southern & Berg, 1982).

SV40 early polyadenylation

signal

Allows efficient transcription termination

and polyadenylation of mRNA.

pUC origin Allows high-copy number replication and

growth in E. coli.

Ampicillin (bla) resistance gene

(β-lactamase)

Allows selection of transformants in E. coli.