pBJ-243 [pRS423-GAL1/10] (ATCC® 77449)

Applications: YE-type (episomal) shuttle vectorYX-type (expression) shuttle vectorexpression vectorshuttle vectorvector containing primer sites useful for sequencingvector permitting RNA synthesis in vitrovector permitting production of single-stranded DNA  /  Depositors: C Hug

Permits and Restrictions

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Designations pBJ-243 [pRS423-GAL1/10]
Depositors C Hug
Biosafety Level 1
Vector Information
Size (kb): 6.4800000190734860
Vector: pBJ-243 (phagemid)
Promoters: Promoter for in vitro transcription T7 (phi10)
Construction: pRS423 (ATCC 77104)
Marker(s):ampR,HIS3
Construct size (kb): 6.480000019073486
Features: marker(s): HIS3
marker(s): ampR
promoter for expression: GAL10...GAL1
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7 (phi10)
replicon: 2 micron
replicon: f1
replicon: pMB1
MCS: BamHI...SacI
MCS: KpnI...EcoRI
Applications
YE-type (episomal) shuttle vector
YX-type (expression) shuttle vector
expression vector
shuttle vector
vector containing primer sites useful for sequencing
vector permitting RNA synthesis in vitro
vector permitting production of single-stranded DNA
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--6.6; EcoRI--6.6; SacI--6.6.
The GAL1 and GAL10 promoters are of similar strength, but the GAL10 promoter has a disrupted RNA start site. Unless a start site is provided by the insert, little or no message should be made from the GAL10 side of the promoter cassette.
Do not use this side of the promoter unless you can insure that mRNA will be properly initiated by providing an RNA start site.
Note that no transcription terminators are provided.
Avoid palindromic restriction enzyme recognition sequences between the transcription start point and the start of translation by placing the AUG of the gene to be expressed as close to the promoter cassette as possible.
Check for inappropriate initiation or termination codons 5' to the cloned insert.
The polylinker sites listed may not be unique in this vector.
YE-type, high copy number, galactose-inducible expression shuttle vector containing a divergent GAL1/GAL10 promoter cassette. The cassette is a 685 bp EcoRI/BamHI fragment with the GAL1 promoter adjacent to the EcoRI site.
Can be used to produce ssDNA. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and yeast REP3 and FRT sequences.
Growth Conditions
Temperature: 37.0°C
References

Christopher Hug, personal communication

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