pHP45omega-Tc [JF297] (ATCC® 37656)

Applications: integrating vectorshuttle vectorvector permitting positive selection for integrationvector useful for interposon mutagenesis  /  Depositors: J Frey

Permits and Restrictions

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Designations pHP45omega-Tc [JF297]
Depositors J Frey
Biosafety Level 1
Vector Information
Size (kb): 4.4800000190734860
Vector: pHP45omega-Tc (plasmid)
Construction: pHP45:omega, pBR322 tetR
Marker(s):ampR,tetR
Construct size (kb): 4.480000019073486
Features: marker(s): ampR, tetR
replicon: pMB1
terminator: T4D gene 32
Applications
integrating vector
shuttle vector
vector permitting positive selection for integration
vector useful for interposon mutagenesis
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--2.5, 2.4; HindIII--2.45, 2.3; SalI--4.9.
Because resistance levels may vary, it is important to determine the proper antibiotic concentration for each strain or isolate.
One of a series (ATCC 37653 through 37657) of vectors for in vitro mutagenesis (via the omega interposon) of gram-negative genera.
pHP45:omega was cut with BssHII + SphI (removing the spcR/strR gene) and the ends were made blunt. This was ligated to a blunt-ended 1.4 kb HindIII/AvaI fragment of pBR322 containing the tetracycline resistance gene.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Fellay R, et al. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of Gram-negative bacteria. Gene 52: 147-154, 1987. PubMed: 3038679

Frey J, Krisch HM. Omega mutagenesis in Gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species. Gene 36: 143-150, 1985. PubMed: 2998930

Shipped freeze-dried
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