HO-poly-KanMX4-HO vector (V009918)

Basic Vector Information

      • Vector Name:
      • HO-poly-KanMX4-HO
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6062 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Voth WP, Richards JD, Shaw JM, Stillman DJ.

HO-poly-KanMX4-HO vector Vector Map

HO-poly-KanMX4-HO6062 bp30060090012001500180021002400270030003300360039004200450048005100540057006000vector 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 KanMX selectable marker; HO region is bracketed by two NotI sites, with polylinker sites between the HO and KanMX4regions

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-poly-KanMX4-HO vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6062 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector HO-poly-KanMX4-HO, complete sequence.
ACCESSION   AF324728
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  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 6062)
  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 6062)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6062
                     /organism="Cloning vector HO-poly-KanMX4-HO"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:153593"
     misc_feature    1..6062
                     /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 KanMX selectable marker; HO region is bracketed by two 
                     NotI sites, with polylinker sites between the HO and KanMX4
                     regions"
     gene            1069..2425
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1069..1412
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1413..2222
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2228..2425
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     2995..3011
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3026..3044)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3051..3067)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3939..4043
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4044..4904
                     /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      5075..5663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5951..5972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    6027..6043
                     /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     6051..5
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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