| Vector |
Construct size (kb): 6.0 DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pCR2.1-TOPO
Intact vector size: 3.908
Type of vector: plasmid
Cloning sites: HindIII KpnI SacI BamHI SpeI BstXI EcoRI EcoRI EcoRV BstXI
NotI XhoI NsiI XbaI ApaI
Polylinker sites: HindIII KpnI SacI BamHI SpeI BstXI EcoRI EcoRI EcoRV BstXI
NotI XhoI NsiI XbaI ApaI
Features (with orientation and position when available):
insert detection: lacZalpha, ->, 1-571
primer site: M13 reverse priming site, ->, 205-221
MCS: bases, ->, 234-357
promoter: T7, ->, 364-383
primer site: M13 forward (-20) priming site, ->, 391-406
primer site: M13 forward (-40) priming site, ->, 411-426
replicon: f1 origin, ->, 548-962
marker(s): kanR, ->, 1296-2090
marker(s): ampR, ->, 2108-2968
replicon: ColE1 origin, ->, 3113-3786
Cross references: |
| Insert |
DNA: cDNA DESCRIPTION OF INSERT COMPONENT:
Genome: Homo sapiens
Gene name: acetyl-CoA synthetase
Contains complete coding sequence?: Y
Strain: human HeLa
Type of DNA: cDNA
Insert end:
Insert end:
Insert size (kb): 2.1
Cross references: Insert lengths(kb): 2.099999904632568 Gene product: acetyl-CoA synthetase Target Gene: acetyl-CoA synthetase |
| Comments |
Restriction digests of the clone give the following sizes (kb): BamHI--6.0; BamHI/NotI--3.9, 2.1; EcoRI--3.8, 2.0.0.2; HindIII--4.1, 1.5,0.3. The pTOPO-ACS contains a 2.1 kb insert that encodes the full length cDNA for human acetyl CoA synthetase (ACS) (amino acids 1-701). THe plasmid was constructed in the following manner. Polymerase chain reaction was used to amplify the full length human ACS with twelve 5 prime untranslated region (UTR) nucleotides and 29 nucleotides of the 3 prime UTR. The PCR product was ligated directly into pCR 2.1-TOPO (Invitrogen). |
| References |
Luong A, et al. Molecular characterization of human acetyl-CoA synthetase, an enzyme regulated by sterol regulatory element-binding proteins. J. Biol. Chem. 275: 26458-26466, 2000. PubMed: 10843999
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