EDRELAINE A.

FLORES BIO 710: Advanced Biotechnology

Master of Science in Biology Education (MSBioEd)

pBR322 Vector

The p stands for "plasmid," and BR for "Bolivar" and "Rodriguez." pBR322 is 4361

base pairs in length and has two antibiotic resistance genes – the gene bla encoding the

ampicillin resistance (AmpR) protein, and the gene tetA encoding the tetracycline

resistance (TetR) protein.

 pBR322 DNA is a commonly used plasmid cloning vector in E. coli. The molecule is

a double-stranded circle 4,361* base pairs in length. pBR322 contains the genes for

resistance to ampicillin and tetracycline, and can be amplified with chloramphenicol.

The molecular weight is 2.83 x 106 daltons. Sequencing data from Watson

(confirmed at New England Biolabs, Inc.) has shown pBR322 to be 4,361 base pairs,

not 4,363 base pairs as previously reported.

GERVY R. VALEROSO BIO 710: Advanced Biotechnology
Master of Science in Biology Education (MSBioEd)

pGEM-3Z Vector
Description:

The pGEM-3Z Vector is intended for use as a standard cloning vector, as

well as for the highly efficient synthesis of RNA in vitro.

The vector carries the lacZ alpha-peptide and the multiple cloning region

arrangement from pUC18 allowing selection of recombinants by blue/white

screening.

In addition, the vector contains both the SP6 and T7 RNA polymerase

promoters flanking the multiple cloning region.

This arrangement gives rise to a functional α-peptide that is capable of

complementing the product of the lacZΔM15 gene to produce functional β-

galactosidase.

Cells with the genotype, lacZΔM15, and also containing the pGEM®-3Z

Vector will be blue when plated on indicator media containing IPTG and X-Gal.

However, when the lacZ α-peptide sequence is disrupted by cloning into

the pGEM®-3Z multiple cloning region, complementation does not occur and

no β-galactosidase activity is produced.

Therefore, bacterial colonies harboring recombinant pGEM®-3Z Vector

constructs remain white.

The pGEM-3Z and pGEM-4Z Vectors are essentially identical except for

the orientation of the SP6 and T7 promoters.

Specifications:

 Item: pGEM-3Z Vector

 Company: Promega

 Catalog Number: P2151

 Quantity: 20 µg

 Promoters: SP6 and T7 promoters

 MCS: Multiple cloning site provides a selection of restriction sites for

cloning. EcoR I, Sac I, Kpn I, Ava I, Xma I, Sma I, BamH I, Xba I, Sal

I, Acc I, Hinc II, Pst I, Sph I, Hind III

 Vector Name: pGEM®-3Z

 Applications: Cloning and Transcription Vectors

  Host: Bacteria

 Origin of Replication: pUC18

 Resistance: Ampicillin (AmpR)

 Selectable Marker: Blue/white screening for recombinants

 Vector Type: Cloning Vectors

 Vector Size: 2743 bp

Features:

 Blue/White Screening: Allows the easy identification of recombinant

clones.

 Versatile: This vector can be used for standard cloning and in vitro

transcription from SP6 and T7 RNA polymerase promoters flanking

the multiple cloning region.

 Convenient: Multiple cloning site provides a selection of restriction

sites for cloning.

Reference:

Yanisch-Perron, C. et al. (1985) Gene 33, 103–19. 0278VA05_4A-W pGEM®-3Z

Vector GenBank® Accession Number X65304 and vector sequence text file.
https://www.biocompare.com/23848-Vector/177049-pGEMsupx00AEsup3Z-
Vector/