pRS415 with LEU2 marker vector (V003593)

Basic Vector Information

Vector Name:
pRS415 with LEU2 marker
Antibiotic Resistance:
Ampicillin
Length:
6021 bp
Type:
Yeast centromere vector
Replication origin:
ori
Host:
Yeast
Source/Author:
Sikorski RS, Hieter P.
Promoter:
LEU2

pRS415 with LEU2 marker vector Map

pRS415 with LEU2 marker6021 bp30060090012001500180021002400270030003300360039004200450048005100540057006000LEU2LEU2 promoterf1 oriM13 fwdT7 promoterMCST3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterCEN/ARS

pRS415 with LEU2 marker vector Sequence

LOCUS       40924_37918        6021 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast centromere vector pRS415 with LEU2 marker, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6021)
  AUTHORS   Sikorski RS, Hieter P.
  TITLE     A system of shuttle vectors and yeast host strains designed for 
            efficient manipulation of DNA in Saccharomyces cerevisiae
  JOURNAL   Genetics 122 (1), 19-27 (1989)
  PUBMED    2659436
REFERENCE   2  (bases 1 to 6021)
  AUTHORS   Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P.
  TITLE     Multifunctional yeast high-copy-number shuttle vectors
  JOURNAL   Gene 110 (1), 119-122 (1992)
  PUBMED    1544568
REFERENCE   3  (bases 1 to 6021)
  AUTHORS   Stillman DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-1993) David J. Stillman, Dept. of Cellular, Viral 
            and Molecular Biology, University of Utah Medical Center, Salt Lake 
            City, UT 84132 USA
REFERENCE   4  (bases 1 to 6021)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; 
            date: "1989"; volume: "122"; issue: "1"; pages: "19-27"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Gene"; 
            date: "1992"; volume: "110"; issue: "1"; pages: "119-122"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-1993) David J. Stillman, Dept. of Cellular, Viral and 
            Molecular Biology, University of Utah Medical Center, Salt Lake 
            City, UT 84132 USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3100..3118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3127..3234)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3247..3265)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3286..3302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3310..3326)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3334..3364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3379..3400)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4450..5307)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5308..5412)
                     /label=AmpR promoter
     misc_feature    5449..5952
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"

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