pET-His (ATCC® 87036)

Applications: expression vector  /  Depositors: T Hai

Permits and Restrictions

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Designations pET-His
Depositors T Hai
Biosafety Level 1
Host
E. coli HB101
Vector Information
Size (kb): 4.636
Vector: pET-His (plasmid)
Promoters: Promoter T7 (phi10)
Construction: pET3a
Marker(s):ampR
Construct size (kb): 4.636
Features: marker(s): ampR
promoter: T7 (phi10)
replicon: pMB1
MCS: NdeI...BamHI
terminator: phi10
Applications
expression vector
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--4.6; BamHI--4.6; BglI--2.5, 2.1, 0.25.
The plasmid contains the following restriction sites (kb from nt 1): BamHI--4.12; BglI--0.87, 3.19, 3.42; BglII--3.99; EcoRI--4.63; HindIII--4.60; NcoI--4.12; NdeI--4.09; PstI--0.75; PvuI--0.62; PvuII--2.29; SalI--3.71; XbaI--4.05; XhoI--4.11.
Digestion with NcoI, BamHI or XhoI and subsequent filling of ends, allows cloning of blunt-ended fragments with a +0, +1 (C) or +2 (GA) nucleotide reading frame, respectively.
Fusion proteins can be purified using a nickel-chelating column, even under denaturing conditions.
Constructed from pET3a by insertion of a double stranded oligonucleotide linker between the NdeI and BamHI sites, creating three restriction sites and encoding six His residues.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Chen BP, Hai T. Expression vectors for affinity purification and radiolabeling of proteins using Escherichia coli as host. Gene 139: 73-75, 1994. PubMed: 8112591

Shipped freeze-dried
Shipping Information

 Freeze dried E. coli containing the plasmid