pGEM-7Zf(+)/LIC-R (ATCC® 87049)

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

Designations pGEM-7Zf(+)/LIC-R
Depositors RS Haun
Biosafety Level 1
Vector Information
Size (kb): 3.0329999923706060
Vector: pGEM-7Zf(+)/LIC-R (phagemid)
Promoters: Promoter T7
Construction: pGEM-7zf(+)
Marker(s):ampR
Construct size (kb): 3.032999992370606
Features: insert detection: lacZ
marker(s): ampR
promoter: SP6
promoter: T7
promoter: lac
replicon: f1
replicon: pMB1
MCS: ApaI...NsiI
Applications
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--3.05; HindIII--3.05; ApaI--3.05.
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 pBluescript II KS(+)/LIC and pGEM-7Zf(+)/LIC-R vectors (ATCC 87047 and 87049).
Differs from pGEM-7Zf(+)/LIC-F (ATCC 87048) only in the orientation of comlementary ends generated at the cloning site.
Ligation-independent cloning vector.
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 freeze-dried
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