pECV25 (ATCC® 77187)

Applications: contains sequence nuclear antigen 1expression vectorshuttle vector  /  Depositors: C Backendorf

Permits and Restrictions

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Designations pECV25
Depositors C Backendorf
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Vector Information
Size (kb): 11.3170003890991200
Vector: pECV25 (plasmid)
Promoters: Promoter RSV LTR
Construction: pRSVneo, p205
Marker(s):G418R,ampR,hygR
Construct size (kb): 11.31700038909912
Features: marker(s): ampR, hygR, G418R
promoter: RSV LTR
replicon: pMB1, EBV oriP
terminator: beta-globin polyadenylation
Applications
contains sequence nuclear antigen 1
expression vector
shuttle vector
Comments
Restriction digests of the clone give the following sizes (kb): HindIII/SacI--5.7 (doublet); SalI--11.5; XhoI--11.5.
Shuttle expression vector maintained episomally in human cells at a copy number of 25 - 50. Shows high transfection efficiency and can be introduced as supercoiled plasmids.
Vector can be rescued from transfected cells with no detectable rearrangements.
The EBNA-1 gene contains an in-frame deletion of a repetitive region of 700 +/- 20 bp. This deletion increases the number of stably maintained plasmid molecules per cell.
The order of the major features in this plasmid is: bla (ampR) - hph (hygR) - RSV LTR - HindIII - rabbit beta-globin intron splicing signals - NotI - agpt (G418R) - XhoI - rabbit beta-globin polyadenylation signal - SalI - oriP - SacI - EBNA-1 - ClaI.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Belt PB, et al. Efficient cDNA cloning by direct phenotypic correction of a mutant human cell line (HPRT-) using an Epstein-Barr virus-derived cDNA expression vector. Nucleic Acids Res. 19: 4861-4866, 1991. PubMed: 1656380

Belt PB, et al. Induction of a mutant phenotype in human repair proficient cells after overexpression of a mutated human DNA repair gene. Nucleic Acids Res. 19: 5633-5637, 1991. PubMed: 1945841

Belt PB, et al. Construction and properties of an Epstein-Barr-virus-derived cDNA expression vector for human cells. Gene 84: 407-417, 1989. PubMed: 2482230