pIIIEx424 tRNA (ATCC® 87175)

Applications: YE-type (episomal) shuttle vectorvector permitting RNA synthesis in vitrovector permitting expression of small structured RNAsvector permitting production of single-stranded DNAvector permitting visual detection of recombinants  /  Depositors: DR Engelke

Permits and Restrictions

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Designations pIIIEx424 tRNA
Depositors DR Engelke
Biosafety Level 1
Host
Distribution host: Escherichia coli HB101 (ATCC 33694)
Vector Information
Size (kb): 7.3000001907348630
DESCRIPTION OF VECTOR:
Intact vector size: 7.300
Type of vector: plasmid
Cloning sites: EcoRI
Polylinker sites:
Construction: pRS424
Host range: Saccharomyces cerevisiae; Escherichia coli
Features (with orientation and position when available):
marker(s): TRP1, ->, 468-1142
replicon: f1, ->, 1241-1707
insert detection: lacZ', <-
promoter for in vitro transcription: T7, ->, 1867-1886
restriction site: SalI
terminator: RPR1, <-
restriction site: EcoRI
promoter: SUP4 tRNA intragenic promoter, <-
restriction site: BamHI
promoter for in vitro transcription: T3, <-
promoter: lac, <-
replicon: pMB1
marker(s): ampR, <-
replicon: 2 micron
Vector: pIIIEx424 tRNA (plasmid)
Promoters: Promoter for in vitro transcription T7
Construction: pRS424
Marker(s):TRP1,ampR
Construct size (kb): 7.300000190734863
Features: insert detection: lacZ'
marker(s): TRP1
marker(s): ampR
promoter: SUP4 tRNA intragenic promoter
promoter: lac
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
replicon: 2 micron
replicon: f1
replicon: pMB1
restriction site: BamHI
restriction site: EcoRI
restriction site: SalI
terminator: RPR1
Applications
YE-type (episomal) shuttle vector
vector permitting RNA synthesis in vitro
vector permitting expression of small structured RNAs
vector permitting production of single-stranded DNA
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--7.4; BamHI--7.4; HindIII--5.2, 2.1.
Cloned inserts can be transcribed from the SUP4 tRNA gene intragenic promoter to produce structured pre-tRNA leaders fused to RNA corresponding to inserted sequences.
One of a series of shuttle vectors (ATCC 87167 - 87182) designed for stable expression of small structured RNAs in yeast. The vectors differ in promoter/terminator cassettes, selectable markers, replicons, and copy number.
Constructed by insertion of a promoter/terminator cassette into the multiple cloning site of the shuttle vector pRS424.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Good PD, Engelke DR. Yeast expression vectors using RNA polymerase III promoters. Gene 151: 209-214, 1994. PubMed: 7828876

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