pNH218 (ATCC® 68103)

Applications: contains sequence alkaline phosphataseexpression vectorproduces protein alkaline phosphataseshuttle vectorvector for other usesvector permitting construction of fusion proteinsvector permitting positive selection for insertsvector with sequences for protein export  /  Depositors: OmniGene, Inc., JG Pero, OmniGene, Inc.

Designations pNH218
Permits and Restrictions

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Depositors OmniGene, Inc., JG Pero, OmniGene, Inc.
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. This material may not have been produced or characterized by ATCC.
Biosafety Level 1
Vector Information
Construct size (kb): 11.5
contains sequence alkaline phosphatase
expression vector
produces protein alkaline phosphatase
shuttle vector
vector for other uses
vector permitting construction of fusion proteins
vector permitting positive selection for inserts
vector with sequences for protein export
Restriction digests of the clone give the following sizes (kb): BamHI--7.8, 3.8; HindIII--6.8, 4.7.
The clone appears to be unstable. We recommend that the ATCC-provided stock be streaked for single colonies and that a colony with the correct restriction pattern be selected.
This culture was previously deposited as ATCC 53063
An Escherichia coli/Bacillus spp. shuttle expression vector for the construction and secretion of fusion proteins. Contains the alkaline phosphatase gene of Escherichia coli.
pBs81/6 (Bacillus replication) was cut with HindIII and cloned into the HindIII site.
A 1.1 kb EcoRI fragment with Bacillus licheniformis alpha-amylase regulatory and signal sequences was cloned into pUC9 cut with EcoRI/SmaI. A 3.0 kb BamHI/HindIII fragment (Escherichia coli phoA minus signal sequences and the first 5-13 aa) was inserted.
Media ATCC Medium 1065 (see below) plus chloramphenicol (5.0 mcg/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0
Growth Conditions
Temperature: 37.0°C

Stephens MA, et al. Secretion vector. US Patent 4,769,327 dated Sep 6 1988

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