YEplac112 vector (V010169)

Basic Vector Information

      • Vector Name:
      • YEplac112
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4989 bp
      • Type:
      • Yeast Plasmids
      • Source/Author:
      • Gietz RD, Sugino A.
      • Copy Number:
      • High copy number
      • 5' Primer:
      • M13 fwd
      • 3' Primer:
      • M13 rev

YEplac112 vector Vector Map

YEplac1124989 bp6001200180024003000360042004800CAP binding sitelac promoterlac operatorM13 revMCSM13 fwd2u oriTRP1TRP1 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 ℃.

YEplac112 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49652        4989 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal plasmid with a TRP1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Gietz RD, Sugino A.
  TITLE     New yeast-Escherichia coli shuttle vectors constructed with in vitro
            mutagenized yeast genes lacking six-base pair restriction sites.
  JOURNAL   Gene 1988;74:527-34.
  PUBMED    3073106
REFERENCE   2  (bases 1 to 4989)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4989)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1988"; volume: "74"; pages: "527-34"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     The sequence of this plasmid was reconstructed using the description
            from Gietz and Sugino.
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(233..289)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(743..2089)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2204..2875)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2876..2977)
                     /label=TRP1 promoter
     promoter        3083..3187
                     /label=AmpR promoter
     CDS             3188..4045
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4219..4807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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