pBluescript II KS(+)/LIC (ATCC® 87047)

Other IDs:

Nucleotide (GenBank) : U25268 Ligation-independent cloning vector pGEM-7Zf(+)/LIC-R, complete sequence.

Nucleotide (GenBank) : U25272 Ligation-independent cloning vector pGEM-7Zf(+)/LIC-F, complete sequence.

 /  Applications: vector permitting visual detection of recombinants  /  Depositors: RS Haun

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Designations pBluescript II KS(+)/LIC
Depositors RS Haun
Other IDs

Nucleotide (GenBank) : U25268 Ligation-independent cloning vector pGEM-7Zf(+)/LIC-R, complete sequence.

Nucleotide (GenBank) : U25272 Ligation-independent cloning vector pGEM-7Zf(+)/LIC-F, complete sequence.

Biosafety Level 1
Vector Information
Size (kb): 3.0000000000000000
Vector: pBluescript II KS(+)/LIC (phagemid)
Promoters: Promoter T3
Construction: pBluescriptIIKS(+)
Marker(s):ampR
Construct size (kb): 3.0
Features: insert detection: lacZ
marker(s): ampR
promoter: T3
promoter: T7 (phi10)
replicon: f1
replicon: pMB1
MCS: KpnI...SacI
Applications
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): KpnI--3.0; XbaI--3.0; BamHI--3.0.
Preparation of the vector for cloning includes linearization with NarI, gel purification of the linearized vector, and treatment with T4 DNA polymerase in the presence of dATP.
The target sequence can be amplified using sequence specific primers modified at the 5' end to contain an additional 13 nt complementary to the vector sequence.
The forward primer should contain 5'-CTGGTTCCGGCGA-3' followed by 12-15 nt target specific sequence. The reverse primer should contain 5'-CTCGCTCCGGCGA-3' followed by 12-15 nt target specific sequence.
Following amplification, the amplified sequence should also be gel purified and treated with T4 DNA polymerase in the presence of dTTP. Annealing of the vector and amplification product forms a duplex molecule that can be used directly for transformation.
Sequences amplified using these primers are also compatible with the pGEM-7Zf(+)/LIC-F and pGEM-7Zf(+)/LIC-R vectors (ATCC 87048 and 87049).
Ligation-independent cloning vector.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Haun RS, et al. Rapid, reliable ligation-independent cloning of PCR products using modified plasmid vectors. BioTechniques 13: 515-518, 1992. PubMed: 1362067

Shipped frozen
Shipping Information

 Frozen glycerol stock of  E. coli containing the phagemid.

Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 87047