| Vector Information |
Size (kb): 5.8000001907348630 DESCRIPTION OF VECTOR:
Intact vector size: 5.800
Type of vector: phagemid
Cloning sites: XhoI SalI EcoRI SmaI BamHI NotI SacI
Polylinker sites: KpnI ApaI XhoI SalI ClaI HindIII EcoRV EcoRI PstI SmaI BamHI
SpeI XbaI EagI NotI BstXI SacII SacI
Construction: pRSS56 [pBluescript KS+, pBS(+)], GAL1/10 promoter
Host range: Candida robustaSaccharomyces cerevisiae; Escherichia coli
Features (with orientation and position when available):
insert detection: lacZ'
marker(s): ampR, URA3
promoter: lac, T3, T7, GAL1
replicon: pMB1, f1, ARS1 Vector: pRS169 (phagemid) Promoters: Promoter T3 Construction: pRSS56 [pBluescript KS+, pBS(+)], GAL1/10 promoter Marker(s):URA3,ampR Construct size (kb): 5.800000190734863 Features: insert detection: lacZ'
marker(s): ampR, URA3
promoter: lac, T3, T7, GAL1
replicon: pMB1, f1, ARS1 |
| Applications |
YC-type (centromeric) 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 vector permitting visual detection of recombinants |
| Comments |
Restriction digests of the clone give the following sizes (kb): PvuI--2.9 (doublet); BamHI--5.8; EcoRI--5.8; EcoRV--4.5, 1.3; HindIII--5.8; SalI--5.8; SmaI--5.8; XbaI--uncut. Expression is inducible by galactose. YX-type expression shuttle vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis and encodes the lacZ alpha (lacZ') peptide. Transcription from the GAL1 promoter (no ATG included in the sequence) proceeds through the polylinker. pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+). A fragment containing the URA3 gene was inserted into the NdeI site and a DraIII/EcoRI fragment containing the GAL1 and GAL10 promoters was inserted into the Asp718 site of the MCS of pRSS56. All ends were blunted. The order of the major features in this plasmid is: URA3 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - GAL1 promoter - GAL10 promoter - pMB1 ori - bla - CEN - ARS1. |
