YEp85 vector (V001489)

Basic Vector Information

      • Vector Name:
      • YEp85
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6801 bp
      • Type:
      • Episomal vector
      • Source/Author:
      • Baruffini E, Serafini F, Lodi T.

YEp85 vector Vector Map

YEp856801 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600CAP binding sitelac promoterlac operatorM13 rev2 micron originTEF-promoterTEF-terminatorAmpR 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 ℃.

YEp85 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6801 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Episomal vector YEp85, complete sequence.
ACCESSION   FJ834448
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Construction and characterization of centromeric, episomal and 
            GFP-containing vectors for Saccharomyces cerevisiae prototrophic 
            strains
  JOURNAL   J. Biotechnol. 143 (4), 247-254 (2009)
  PUBMED    19683551
REFERENCE   2  (bases 1 to 6801)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2009) Department of Genetics, Biology of 
            Microorganisms, Anthropology, Evolution, University of Parma, Viale 
            Usberti 11/A, Parma 43100, Italy
REFERENCE   3  (bases 1 to 6801)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6801
                     /organism="Episomal vector YEp85"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:670503"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     CDS             216..530
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIYRIVLVTHH
                     "
     misc_feature    233..289
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    244..285
                     /note="multiple cloning site; MCS"
     misc_feature    288..529
                     /note="beta-galactosidase alpha fragment; LacZ'"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      740..1905
                     /label=2 micron origin
                     /note="2 micron origin"
     rep_origin      743..2089
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     regulatory      2196..2586
                     /regulatory_class="promoter"
                     /note="TEF-promoter"
     gene            2243..3599
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2243..2586
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2587..3396
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="aminoglycoside 3'-phosphotransferase; APH"
                     /protein_id="ACV50013.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2587..3396
                     /gene="kanR"
                     /label=kanR
     regulatory      3394..3628
                     /regulatory_class="terminator"
                     /note="TEF-terminator"
     terminator      3402..3599
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4895..4999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5000..5860
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase"
                     /protein_id="ACV50014.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5000..5860
                     /gene="ampR"
                     /label=ampR
     rep_origin      6031..6619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

This page is informational only.