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pTH4 [TRP1 --> HIS3 converter]
pTH4 [TRP1 --> HIS3 converter]
規(guī)格:
貨期:
編號(hào):B228401
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 pTH4 [TRP1 --> HIS3 converter]
商品貨號(hào) B228401
Designations pTH4 [TRP1 --> HIS3 converter]
Depositors FR Cross
Biosafety Level 1
Vector Information
Size (kb): 6.7
Vector: pTH4 (phagemid)
Construction: pBC KS+
Marker(s): cmlR, kanR, HIS3
gene disruption cassette: tr:HIS3/kanR
replicon: f1
replicon: pMB1
restriction site: SmaI, EcoRI
restriction site: XhoI
Applications
YI-type (integrating) shuttle vector
host modification
marker swap vector TRP1 --> HIS3
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--6.7; HindIII--4.7, 1.8, 0.2; EcoRI/XhoI--3.5, 3.3.
To convert the host phenotype from TRP1 to HIS3, transform with the XhoI+EcoRI digested vector and select for His+ transformants.
Some combinations of marker swap plasmids and target locus may result in relatively high reversion rates. In most but not all cases the frequencies of successful convertants are greater than 30%.
A marker swap vector designed to change the S. cerevisiae host phenotype by one-step gene disruption of the TRP1 gene with the HIS3 and kanR markers.
When swapping markers on an episomal plasmid, appropriate phenotype may result from loss of the plasmid unless a second selectable or scorable marker is used to ensure plasmid maintenance.
Vector was constructed by replacing an internal EcoRV fragment of TRP1 with a SmaI fragment containing the HIS3 and kanR coding sequences. HIS3 and TRP1 are in the same orientation.
Media ATCC® Medium 2994: Medium 1065 plus Kanamycin 50μg/ml plus Chloramphenicol 25μg/ml
Growth Conditions
Temperature: 37°C
References

Cross FR. 'Marker swap' plasmids: convenient tools for budding yeast molecular genetics. Yeast 13: 647-653, 1997. PubMed: 9200814

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