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yGALset351 vector (V001482)

Basic Vector Information

Vector Name:
yGALset351
Antibiotic Resistance:
Ampicillin
Length:
6852 bp
Type:
Cloning vector
Replication origin:
ori
Host:
Yeast
Source/Author:
Enomoto S, Chen G, Berman J.
Promoter:
GAL1,10

yGALset351 vector Vector Map

yGALset3516852 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600T7 promoterGAL1,10 promoterRBS6xHisT7 tag (gene 10 leader)Xpress(TM) tagmultiple cloning sitesT7 terminatorCEN/ARSLEU2LEU2 promoterAmpR promoterAmpRori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

yGALset351 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49697        6852 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset351, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  AUTHORS   Enomoto S, Chen G, Berman J.
  TITLE     Vectors for expressing T7 epitope- and His6 affinity-tagged fusion 
            proteins in S. cerevisiae
  JOURNAL   BioTechniques 24 (5), 782-786 (1998)
  PUBMED    9591127
REFERENCE   2  (bases 1 to 6852)
  AUTHORS   Enomoto S, Berman J.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-1998) Plant Biology, University of Minnesota, 220 
            Biological Sciences Center, St. Paul, MN 55108, USA
REFERENCE   3  (bases 1 to 6852)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6852)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "24"; issue: "5"; pages: 
            "782-786"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-JAN-1998) Plant Biology, University of Minnesota, 220 Biological
            Sciences Center, St. Paul, MN 55108, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..88
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(121..785)
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     RBS             804..826
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             846..863
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             867..899
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             903..926
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     misc_feature    934..980
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     terminator      1044..1091
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    complement(1805..2967)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     CDS             complement(3332..4423)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4436..4840)
                     /label=LEU2 promoter
     promoter        4946..5050
                     /label=AmpR promoter
     CDS             5051..5908
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6082..6670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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