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p3PK
p3PK
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編號:B233968
品牌:Mingzhoubio

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產(chǎn)品名稱 p3PK
商品貨號 B233968
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

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