PureBiotech LLC Online - LabLink - pGATA Positive Selection Vector

Index: Volume 1 Number 1
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NO BACKGROUND cloning
Eliminate the colony screening step - all colonies are positive
Works with any strain of E. coli
No need for dephosphorylation or complete digestion of vector
Quick and easy cloning - sure to work every time

Figure 1: Map of the CloneSure™ pGATA Positive Selection Vector

The CloneSure™ pGATA positive selection vector is a general cloning vector which contains a gene coding for a toxic peptide that inhibits bacterial growth when expression is induced. Insertion of a DNA fragment into the multiple cloning site disrupts the gene coding for the toxic peptide, thus rendering the recombinants non-toxic. The vector, in the absence of the insert, produces a toxic peptide which inhibits bacterial replication. Therefore, only colonies which harbor a plasmid containing your cloned insert will grow. This constitutes a no background positive selection system for recombinant molecules.

The toxic peptide in CloneSure™, which represents a region of the eukaryotic transcription factor GATA-1 that binds to the bacterial origin of replication, is under the control of the IPTG-inducible tac promoter. To prevent leaky expression of the fusion protein in the absence of IPTG, the CloneSure™ vector also contains the lacI^q repressor gene. The multiple cloning site (see Figure 1 on page 3) is flanked by T3 and T7 RNA polymerase promoters to allow for in vitro transcription of your cloned insert. Additionally, CloneSure™ contains the ampicillin resistance gene and a high copy number origin of replication (colEI type). Unlike other similar vectors which require special strains of E. coli to accomplish no background cloning, CloneSure™ can be used effectively with any strain of E. coli.

Next Article: From Cells to RT-PCR Results in 6 Hours