Is pBluescript a restriction enzyme?

The pBluescript II SK+ phagemid has an extensive polylinker with 21 unique restriction enzyme recognition sites. It is a high copy number ColE1-based phagemid with a large and versatile polylinker Sac–>Kpn; f1 origin in (+) orientation. The T3 and T7 promoters are for in vitro transcription of RNA.

Why is pBluescript useful?

The versatility of pBluescript vectors alleviates the need for successive subcloning. With 21 restriction sites, in two orientations, most cloned genes can be inserted directionally into the pBluescript polylinkers. If a new gene of unknown sequence is to be cloned, libraries will need to be generated for screening.

Which type of vector is pBluescript?

The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.

What is the size of pBluescript?

Plasmid: pBluescript SK (+)

What is lambda phage vector?

Enterobacteria phage λ (lambda phage, coliphage λ) is a bacterial virus, or bacteriophage, that infects the bacterial species Escherichia coli. Lambda phage consists of a virus particle including a head (also known as a capsid), tail and tail fibers. The head contains the phage’s double-stranded circular DNA genome.

What is polylinker sequence?

A polylinker is a short DNA sequence containing two or more different sites for cleavage by restriction enzymes. Polylinkers are introduced into vectors to make cloning easier by providing sites that allow cloning DNA, cut with any of a number of different restriction enzymes, into a single plasmid.

What is phagemid vector?

A phagemid or phasmid is a DNA-based cloning vector, which has both bacteriophage and plasmid properties. These vectors carry, in addition to the origin of plasmid replication, an origin of replication derived from bacteriophage.

Why is pUC19 a good cloning vector?

pUC19 is a commonly used cloning vector that conveys the Amp resistance. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. NEB offers a selection of common cloning plasmids and DNAs for use as substrates.

Is Agrobacterium a cloning vector?

What is an Agrobacterium vector? In cloning, a vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell, where it can be replicated and expressed. The Agrobacterium tumor-inducing plasmid or Ti-plasmid does the DNA transfer.

What are the advantages of lambda phage vector?

The efficiency of packaging and infection, and the simplicity of plaque screening are advantages of lambda as a cloning vector. A number of ingenious modifications help overcome the disadvantages associated with its mode of growth and its size.

What are the 2 most commonly used vectors?

Two types of vectors are most commonly used: E. coli plasmid vectors and bacteriophage λ vectors. Plasmid vectors replicate along with their host cells, while λ vectors replicate as lytic viruses, killing the host cell and packaging the DNA into virions (Chapter 6).

What happens if insert DNA is cut with two different restriction enzymes at the ends?

What happens if insert DNA is cut with two different restriction enzymes at the ends? Explanation: If the DNA is cut with two different enzymes at the ends, it is possible to ligate the fragment in only one orientation. It is so because each end would have a unique sequence to ligate.

What can a pBluescript 2 SK + vector be used for?

pBluescript II SK+. The pBluescript II SK+ phagemid (plasmids with a phage origin) is a cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.

What kind of promoter is used in pBluescript 2?

Flanking the polylinker are T7 and T3 RNA polymerase promoters that can be used to synthesize RNA in vitro.1, 2The choice of promoter used to initiate transcription determines which strand of the insert cloned into the polylinker will be transcribed. Circular maps and lists of features for the pBluescript II phagemids are shown in figures 1 and 2.

Which is the correct orientation for pBluescript 2?

pBluescript II (+) and (–) are available with two polylinker orientations designated as either KS or SK using the following convention: (1) in the KS orientation, the Kpn I restriction site is nearest the lacZ promoter and the

Which is the reversed MCs in pBluescript II?

Standard cloning vector (phagemid excised from lambda ZAPII). The f1 (–) orientation allows rescue of antisense strand ssDNA. pBluescript II KS (–) has a reversed MCS. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer.