YCp50-poly vector (V010196)

Basic Vector Information

      • Vector Name:
      • YCp50-poly
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7060 bp
      • Type:
      • Yeast Plasmids
      • Copy Number:
      • High copy number
      • 3' Primer:
      • M13 rev

YCp50-poly vector Vector Map

YCp50-poly7060 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900MCSM13 revlac operatorlac promoterURA3 promoterURA3CEN/ARSoriAmpRAmpR promoter

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 ℃.

YCp50-poly vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7060 bp ds-DNA    circular SYN 28-2-2015
DEFINITION  Derivative of yeast centromeric vector YCp50 containing the pUC19 
            polylinker.
ACCESSION   .
VERSION     .
KEYWORDS    YCp50-poly
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7060)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     YCp50-poly was constructed in two steps: (1) YCp50 was cleaved with 
            XhoI and Asp718, blunted and ligated. (2) This construct was then 
            cleaved with EcoRI and SalI, and the EcoRI-PvuII fragment of pUC19 
            was inserted.
FEATURES             Location/Qualifiers
     source          1..7060
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     misc_feature    1..57
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(70..86)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    94..110
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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(118..148)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        1026..1241
                     /gene="S. cerevisiae URA3"
                     /note="URA3 promoter"
     CDS             1242..2045
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /note="URA3"
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    3390..4768
                     /note="CEN/ARS"
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      complement(5236..5824)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5995..6855)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6856..6960)
                     /gene="bla"
                     /note="AmpR promoter"

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