HO-hisG-URA3-hisG-poly-HO vector (V009920)

Basic Vector Information

      • Vector Name:
      • HO-hisG-URA3-hisG-poly-HO
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 8487 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Voth WP, Richards JD, Shaw JM, Stillman DJ.

HO-hisG-URA3-hisG-poly-HO vector Vector Map

HO-hisG-URA3-hisG-poly-HO8487 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400vector designed for targeted integration to the HO locus in yeast; plasmid contains two fragments from the HO promoter inserted in the pUC21-NotI polylinker, along with the hisG-URA3-hisG selectable marker; HO region is bracketed by two NotI sites, with polylinker sites between the HO and hisG-URA3-hisG regions

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 ℃.

HO-hisG-URA3-hisG-poly-HO vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8487 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector HO-hisG-URA3-hisG-poly-HO, complete sequence.
ACCESSION   AF324729
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Voth WP, Richards JD, Shaw JM, Stillman DJ.
  TITLE     Yeast vectors for integration at the HO locus
  JOURNAL   Nucleic Acids Res. 29 (12), E59 (2001)
  PUBMED    11410682
REFERENCE   2  (bases 1 to 8487)
  AUTHORS   Voth WP, Richards J, Shaw J, Stillman DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-NOV-2000) Pathology, University of Utah, 50 N. Medical
            Drive, 5C124 SOM, Salt Lake City, UT 84132, USA
REFERENCE   3  (bases 1 to 8487)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8487
                     /organism="Cloning vector HO-hisG-URA3-hisG-poly-HO"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:153591"
     misc_feature    1..8487
                     /note="vector designed for targeted integration to the HO 
                     locus in yeast; plasmid contains two fragments from the HO 
                     promoter inserted in the pUC21-NotI polylinker, along with 
                     the hisG-URA3-hisG selectable marker; HO region is 
                     bracketed by two NotI sites, with polylinker sites between 
                     the HO and hisG-URA3-hisG regions"
     promoter        2199..2414
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2415..3218
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     5420..5436
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5451..5469)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5476..5492)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6364..6468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6469..7329
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7500..8088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8376..8397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    8452..8468
                     /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     8476..5
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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