LOCUS       Exported                4303 bp ds-DNA    circular SYN 29-11-2013
DEFINITION  Yeast/E. coli integrating shuttle vector with a URA3 marker.
ACCESSION   L14759
VERSION     .
KEYWORDS    YIp352
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4303)
  AUTHORS   Hill JE, Myers AM, Koerner TJ, Tzagoloff A.
  TITLE     Yeast/E. coli shuttle vectors with multiple unique restriction 
            sites.
  JOURNAL   Yeast 1986;2:163-7.
  PUBMED    3333305
REFERENCE   2  (bases 1 to 4303)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4303
                     /organism="Shuttle vector YIp352"
                     /focus
                     /mol_type="genomic DNA"
                     /db_xref="taxon:118282"
     source          1..285
                     /organism="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:562"
     source          286..1610
                     /organism="Saccharomyces cerevisiae"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:4932"
     source          1611..4303
                     /organism="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:562"
     CDS             1..291
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIG"
     misc_feature    15..71
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(75..91)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(364..1167)
                     /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"
     promoter        complement(1168..1378)
                     /gene="S. cerevisiae URA3"
                     /note="URA3 promoter"
     promoter        2182..2286
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             2287..3147
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3318..3906
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4230..4260
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4268..4284
                     /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)."
     primer_bind     4292..5
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"