HO-poly-KanMX4-HO vector

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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
  JOURNAL   Exported Dec 18, 2018 from SnapGene 4.1.8
            http://www.snapgene.com
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"