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| 产品名称： | p3PK |
|---|---|
| 商品货号： | TS142949 |
| Designations： | p3PK |
| Depositors： | JV Frangioni, BG Neel |
| Biosafety Level： | 1 |
| Host： | Distribution host: Escherichia coli DH5alpha |
| Vector Information： | Size (kb): 4.9000000953674320 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pJ3omega Intact vector size: 3.500 Type of vector: plasmid Cloning sites: HindIII PvuI SalI XbaI BamHI SmaI SstI EcoRI ClaI KpnI BglII Polylinker sites: HindIII PvuI SalI XbaI BamHI SmaI SstI EcoRI ClaI KpnI BglII Construction: pBR322, SV40 Host range: mammalian cells; Escherichia coli Features (with orientation and position when available): promoter: SV40 early, -> replicon: SV40 MCS: HindIII...BglII, -> other: SV40 t IVS, -> terminator: SV40, -> marker(s): ampR, -> replicon: pMB1 Cross references: Vector: p3PK (plasmid) Promoters: Promoter SV40 early Construction: pJ3omega, PKM2 Marker(s):ampR Construct size (kb): 4.900000095367432 Features: marker(s): ampR promoter: SV40 early replicon: pMB1, SV40 terminator: SV40 |
| Applications： | contains sequence pyruvate kinase, muscle expression vector in another host, produces protein pyruvate kinase, muscle shuttle vector |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--4.9; BglII--4.9; XbaI--4.9; BamHI--4.9; EcoRI/HindIII--3.4, 1.5; EcoRI/BglII--3.4, 1.5. A shuttle expression vector permitting rapid cloning of putative intracellular targeting sequences as N-terminal, C-terminal, and/or internal fusions with the normally cytosolic chicken muscle pyruvate kinase (PKM2). Permits one step subcloning into any of the pJ(X)omega mammalian expression vectors (ATCC 37719-37724) for versatile promoter control, and into the pGE(X) bacterial fusion protein purification system. The PKM2 sequence encodes amino acids 17-476. The PKM2 coding region contains a unique SalI site. The BamHI site consists of 2 engineered codons added to the end of the PKM2 sequence and immediately followed by an in-frame TGA stop codon. The other 3 sites are outside the reading frame. The order of the major features in the plasmid is: pMB1 ori - SV40 ori - SV40 early promoter - HindIII - Kozak/ translation start sequence - BglII - PKM2 sequences - translation stop - BamHI - XbaI - EcoRI - ClaI - SV40 polyadenylation - ampR. |
| Media： | ATCC® Medium 1946: Terrific broth (ATCC medium 1743) with 100 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Frangioni JV, Neel BG. Use of a general purpose mammalian expression vector for studying intracellular protein targeting: identification of critical residues in the nuclear lamin A/C nuclear localization signal. J. Cell Sci. 105: 481-488, 1993. PubMed: 8408279 John V Frangioni, personal communication |
| Shipped： | freeze-dried |
| Shipping Information： | Distributed: freeze-dried |