pADH59 (ATCC® 39335)

Organism: Homo sapiens, human  /  Clone Type: Clone  /  Depositors: Amgen, Inc., DL Hare, Amgen, Inc.

Permits and Restrictions

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Designations pADH59
Species Homo sapiens, human
Depositors Amgen, Inc., DL Hare, Amgen, Inc.
Applications
produces protein epidermal growth factor urogastrone
Vector
DESCRIPTION OF VECTOR COMPONENT:
Name of vector: unknown
Intact vector size:
Type of vector: plasmid
Vector end: EcoRI
Vector end: SalI
Cloning sites:
Polylinker sites:
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): ampR
Cross references:
Insert
DNA: Synthetic
DESCRIPTION OF INSERT COMPONENT:
Genome: human
Gene symbol: EGF
Genomic copy number: unique
Gene name: epidermal growth factor
Contains complete coding sequence?: U
Chromosome: 4; Localization: 4 q25
Type of DNA: synthetic
Insert 5' end: EcoRI
Insert 3' end: SalI
Insert size (kb): 1.75
Cross references:
Insert lengths(kb): 1.75
Gene product: epidermal growth factor(urogastrone) [EGF]
Alleles: A1, B1, B2, A2
Target Gene: epidermal growth factor
Insert Size (kb) 1.75
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1
Shipping Information Distributed: freeze-dried
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI/SalI--3.3, 1.75; EcoRI--5.1; SalI--5.1; BamHI--4.9, 0.2.
Runaway vector contains a temperature-sensitive mutation in the copy-control gene. Maintained at a low copy number below 34C; replication increases 50-fold at temperatures greater than 34C. Growth at 37C is lethal.
The 1.75 kb EcoRI/SalI fragment contains a trp promoter/regulator and the EGF/urogastrone coding region. The complete coding sequence of the mature peptide, plus a "pro" sequence of 8 aa, is contained in a 0.2 kb XbaI/SalI fragment.
Directs the synthesis of a peptide of 61 aa, including an 8-residue "leader" sequence.
Expresses the urogastrone analog product on the order of 50 mg per O. D. liter as determined by SDS-PAGE. The microbially expressed product is biologically active.
Constructed from oligonucleotides selected to reflect the codon usage of Escherichia coli. Contains a HinfI site interrupting Asp3, an SphI site interrupting Cys31, and an NruI site after Tyr44.
References

Banks AR, Hare DL. Manufacture and expression of genes for urogastrone and polypeptide analogs thereof. US Patent 4,764,593 dated Aug 16 1988

Product Sheet
Product Sheet
Product Sheet