pFC54.t (ATCC® 39789)

Applications: cloning vectorexpression vector  /  Depositors: Cetus Corp., A Belt, Cetus Corp.

Permits and Restrictions

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Designations pFC54.t
U.S. Patent
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished.
Depositors Cetus Corp., A Belt, Cetus Corp.
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Host
Distribution host: Escherichia coli
Vector Information
Size (kb): 5.6100001335144040
DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pFC54
Intact vector size: 5.440
Type of vector: plasmid
Cloning sites:
Polylinker sites:
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): ampR
promoter: lambdaPL
replicon: pMB1
Cross references:
Vector: pFC54.t (plasmid)
Construction: pFC54, pHCW801
Marker(s):ampR
Construct size (kb): 6.099999904632568
Applications
cloning vector
expression vector
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--5.0, 1.15; BamHI--6.1; HindIII--6.1; HindIII/BamHI--5.6, 0.5.
Construct contains a positive retroregulatory element which increases the level of production of the IL2 (Ser125) using a temperature regulated control cassette in a temperature-regulated ColE1 copy mutant plasmid vector.
This should not be grown at temperatures exceeding 30C. The depositor recommends using ampicillin at 100 mcg/ml. The strain is temperature-inducible.
The clone contains a lambdaPL promoter.
This was constructed from a SphI/XbaI fragment of pFC54 ligated to a 655 bp SphI/XbaI fragment of pHCW801 containing the C-terminal 255 bp of Ser125 IL-2, 36 bp of 3' untranslated IL-2 cDNA and 394 bp of the positive retroregulatory element.
Media ATCC® Medium 1315: LB Medium (ATCC medium 1065) with 100 mg/L ampicillin
Growth Conditions
Temperature: 30.0°C
References

Mark DF, et al. Human tumor necrosis factor. US Patent 4,677,063 dated Jun 30 1987

Mark DF, et al. Human tumor necrosis factor. US Patent 4,677,064 dated Jun 30 1987

Gelfand DH, et al. Polypeptide expression using a portable temperature sensitive control cassette with a positive retroregulatory element. US Patent 4,666,848 dated May 19 1987

Shipped freeze-dried
Shipping Information Distributed: freeze-dried
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