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"