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.

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Designations pNH218
Depositors OmniGene, Inc., JG Pero, OmniGene, Inc.
Biosafety Level 1
Vector Information
Construct size (kb): 11.5
Applications
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
Comments
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
References

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

Shipped freeze-dried
Shipping Information Distributed: freeze-dried
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