The pETBlue™-1 vector is designed to identify recombinants by traditional blue/white screening while also allowing T7
lac promoter based expression of target genes. Screening is independent of expression because the T7
lac expression promoter is in an opposed orientation relative to the
E. coli promoter that mediates blue/white screening. pETBlue-1 facilitates the expression of native unfused proteins and allows convenient subcloning of target genes already fused to existing detection and purification tags. The
EcoR V cloning site is appropriately spaced downstream of an
E. coli ribosome binding site. Inserts must encode an ATG start codon at their 5′ end if expression is desired. The sequence is numbered from the first base of the T7 promoter sequence.
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Product information
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Avoid freeze/thaw
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Yes
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Components
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| 20 rxn | 40 rxn
| | • | 2 × 0.5 µg | 4 × 0.5 µg | Blunt Vector
| | • | 10 µl | 10 µl | Positive Control Insert
| | • | 100 µl | 2 × 100 µl | End Conversion Mix |
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Features and benefits
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- No restriction enzymes or special primers
- Compatible with any DNA polymerase
- Independent of 3′-dA addition or sequence context of DNA ends
- Blue/white screening with seven different vectors, including pETBlue expression vectors
- Simple protocol takes less than one hour* from PCR product to plating transformants
* All times listed refer to kits using NovaBlue Singles™ Competent Cells and ampicillin or chloramphenicol selection. Giga kits, which use NovaBlue GigaSingles™ Competent Cells, require one hour for outgrowth.
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Store and ship information
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Storage
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≤ -70°C
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Ship
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Dry Ice Only
Standard Handling
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