pNKY48 (ATCC® 77182)

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

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Designations pNKY48
Depositors N Kleckner
Biosafety Level 1
Vector Information
Size (kb): 4.3000001907348630
Vector: pNKY48 (plasmid)
Construction: pGC-1, pNKY2029
Marker(s):URA3,ampR
Construct size (kb): 4.300000190734863
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): PstI--4.4; BglII--4.4; HindIII--3.1, 1.3; EcoRI--4.4; 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 by inserting a 1.4 kb HindIII fragment from pNKY2029 containing the HIS4/URA3 fusion into pNKY47, which is a derivative of pGC-1 with the original HindIII site destroyed and a new one created at the BamHI site of the MCS.
The order of the major features in this plasmid is: M13 primer - ClaI - XhoI - SalI - XbaI (stop) - BamHI - HindIII - 90 aa HIS4C bridge/URA3 - BamHI - HindIII - BamHI - 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
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