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| 产品名称： | pSVA3 |
|---|---|
| 商品货号： | TS135065 |
| Designations： | pSVA3 |
| Species： | Cricetulus griseus, hamster, Chinese |
| Depositors： | PW Melera |
| Applications： | in another host, produces protein dihydrofolate reductase |
| Vector： | Construct size (kb): 5.099999904632568 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pSV2-neo Intact vector size: 5.729 Type of vector: plasmid Vector end: HindIII Vector end: SmaI Cloning sites: EcoRI BamHI PvuII PstI HindIII Polylinker sites: Construction: SV40, Tn5, pBR322 Host range: vertebrate cells; Escherichia coli Features (with orientation and position when available): replicon: SV40 marker(s): G418R, restriction site: HindIII promoter: SV40 early, replicon: pMB1 marker(s): ampR, Cross references: DNA Seq. Acc.: U02434 |
| Insert： | DNA: cDNA DESCRIPTION OF INSERT COMPONENT: Genome: hamster, Chinese Gene symbol: DHFR Gene name: dihydrofolate reductase Contains complete coding sequence?: U Tissue: lung cell line Type of DNA: cDNA Insert 5 end: HindIII Insert 3 end: BglII Insert size (kb): 0.71 Cross references: Insert information: Insert size (kb): 0.71 DNAxa0: cDNA Genexa0: DHFR Sourcexa0: hamster, Chinese lung cell line Insert lengths(kb): 0.7099999785423279 Tissue: lung cell line Gene product: dihydrofolate reductase DHFR Target Gene: dihydrofolate reductase |
| Insert Size (kb)： | 0.710 |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| 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. |
| Shipping Information： | Distributed: freeze-dried |
| Comments： | Restriction digests of the clone give the following sizes (kb): BamHI--3.5, 1.7; EcoRI--2.7, 2.3; PstI--1.9, 1.4, 0.95, 0.9; PvuII--4.0, 1.1; HindIII/SmaI--5.2. Transfected cells amplify DHFR sequences. Encodes the L22F mutant of DHFR that can serve as a dominant selection marker because it retains catalytic activity and also confers methotrexate (MTX) resistance at low MTX concentrations (125 nM). Constructed by cloning a HindIII/SmaI fragment containing the gene into pSV2-neo from which neoR had been removed. This plasmid was then cut with SmaI and BglII to remove 18 G nucleotides at the 3 end. The BglII site was filled in, and ligated to the SmaI site, thereby destroying the original BglII and SmaI sites. Removing the HindIII/SmaI fragment from pSV2-neo removes all but the 3 171 bp of neoR. |
| References： | Hussain A, et al. Construction of a dominant selectable marker using a novel dihydrofolate reductase. Gene 112: 179-188, 1992. PubMed: 1555767 |