pNKY70 (ATCC® 77183)

Applications: YI-type (integrating) shuttle vectorcontains easily purifiable cassette(s) for constructioncontains sequence histidinol dehydrogenasecontains sequence uridine monophosphate synthetase UMP synthase, orotate phosphoribosyltransferase, orotidine 5'-phosphate decarboxylase, orotate phosphoribosyltransferase 1vector permitting construction of fusion proteinsvector permitting production of single-stranded DNA  /  Depositors: N Kleckner

Permits and Restrictions

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Designations pNKY70
Depositors N Kleckner
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Vector Information
Size (kb): 4.4000000953674320
Vector: pNKY70 (plasmid)
Construction: pNKY48 (ATCC 77182)
Marker(s):URA3,ampR
Construct size (kb): 4.400000095367432
Features: marker(s): ampR, URA3
replicon: pMB1, M13
terminator: URA3
Applications
YI-type (integrating) shuttle vector
contains easily purifiable cassette(s) for construction
contains sequence histidinol dehydrogenase
contains sequence uridine monophosphate synthetase UMP synthase, orotate phosphoribosyltransferase, orotidine 5'-phosphate decarboxylase, orotate phosphoribosyltransferase 1
vector permitting construction of fusion proteins
vector permitting production of single-stranded DNA
Comments
Restriction digests of the clone give the following sizes (kb): BglII--4.4; EcoRI--2.9, 1.5; PstI--4.4; BamHI--3.2, 1.2; XbaI--4.4.
Contains a HIS4C/URA3 fusion cassette that can be used to make fusions to other proteins of interest. Also includes a M13 sequencing primer approximately 300 bp from the HIS4C/URA3 fusion junction.
Encodes 93 amino acids of HIS4C and the complete 267 amino acids of URA3.
Can be used for promoter/enhancer trap screens.
Constructed from pNKY48 (ATCC 77182) by inserting an EcoRI 8-mer at the HindIII site in the polylinker.
The order of the major features in this plasmid is: M13 primer - ClaI - XhoI - SalI - XbaI (stop) - BamHI - EcoRI - HIS4C bridge/URA3 - BamHI - HindIII - BglII - NcoI - EcoRI - M13 ori - pMB1 ori - ampR.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Alani E, Kleckner N. A new type of fusion analysis applicable to many organisms: protein fusions to the URA3 gene of yeast. Genetics 117: 5-12, 1987. PubMed: 3311876

Riles L, et al. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6-kilobase pairs. Genetics 134: 81-150, 1993. PubMed: 8514151

Nancy Kleckner, personal communication

Shipped freeze-dried
Shipping Information Freeze dried E. coli containing the plasmid.
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