pHK9 (ATCC® 87709)

Applications: cloning vector  /  Depositors: F Spencer

Permits and Restrictions

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Designations pHK9
Depositors F Spencer
Biosafety Level 1
Vector Information
Size (kb): 10.8999996185302700
Vector: pHK9 (plasmid)
Marker(s):ampR
Construct size (kb): 10.89999961853027
Features: marker(s): ampR, LYS2
replicon: ori
MCS: SalI (delta15)
restriction site: EcoRI,BamHI,XhoI,HpaI,NruI,BglII,PstI
restriction site: PvuII
coding sequence: karldelta15
Applications
cloning vector
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--10.9; EcoRI--6.5, 2.4, 2.05; PstI--9.0, 1.8; PvuII--10.9.
Restriction digests of the clone give the following sizes (kb): BglII--7.8; EcoRI--7.8; HindIII--5.2, 2.5; EcoRI/HindIII--5.2, 2.5.
Supports introduction of kar1 delta 15 mutation into lys2 yeast strains by two step gene replacement. The karl delta 15 mutation supports transfer of YAC clones between yeast strains by Karl-mating (REF).
To integrate pHK9, linearize within the kar1 gene at the unique PvuII site. Cells that have lost the LYS2 marker through mitotic excision can be selected on alpha- aminoadipate.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Vallen EA, et al. Separate domains of KAR1 mediate distinct functions in mitosis and nuclear fusion. J. Cell Biol. 117: 1277-1287, 1992. PubMed: 1607389

Spencer F, et al. Yeast kar1 mutants provide an effective method for YAC transfer to new hosts. Genomics 22: 118-126, 1994. PubMed: 7959757

Hugerat Y, et al. A versatile method for efficient YAC transfer between any two strains. Genomics 22: 108-117, 1994. PubMed: 7959756

Spencer F, Simchen GTransfer of YAC clones to new hosts by karyogamy deficient mating.In: Spencer F, Simchen GCurrent Protocols in Human GeneticsNew York, N.Y.John Wiley and Sonsin press, 1995

Shipped freeze-dried
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