pNH218 (ATCC® 53063)

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.

Permits and Restrictions

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Designations pNH218
Depositors OmniGene, Inc., JG Pero, OmniGene, Inc.
Biosafety Level 1
Host
Distribution host: Bacillus subtilis 168
Vector Information
Size (kb): 11.5000000000000000
DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pNH218
Intact vector size: 11.500
Type of vector: plasmid
Cloning sites:
Polylinker sites:
Construction: pUC9, pBs81/6, phoA, alpha-amylase signals
Host range: Bacillus sp.; Escherichia coli
Features (with orientation and position when available):
marker(s): cmlR
promoter: alpha-amylase
replicon: pMB1, pBs81/6
Cross references:
Vector: pNH218 (plasmid)
Promoters: Promoter alpha-amylase
Construction: pUC9, pBs81/6, phoA, alpha-amylase signals
Marker(s):cmlR
Construct size (kb): 11.5
Features: marker(s): cmlR
promoter: alpha-amylase
replicon: pMB1, pBs81/6
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--5.8, 5.8; HindIII--5.4, 4.2, 1.3.
A new deposit was made for this clone. It is recommended that ATCC 68103 be used instead of 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 first 5-13 aa) was inserted.
Media Medium 1065: LB Agar/Broth, Miller
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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