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Distribution host: Escherichia coli DH5alpha

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

contains sequence pyruvate kinase, muscle

expression vector

in another host, produces protein pyruvate kinase, muscle

shuttle vector

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.

Temperature: 37.0°C