LOCUS       Exported               11766 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDE215, complete sequence.
ACCESSION   KF170375
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11766)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van 
            Maris AJ.
  TITLE     Replacement of the Saccharomyces cerevisiae acetyl-CoA synthetases 
            by alternative pathways for cytosolic acetyl-CoA synthesis
  JOURNAL   Metab. Eng. (2013) In press
  PUBMED    24269999
REFERENCE   2  (bases 1 to 11766)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran J-M.G., Pronk JT, 
            van Maris AJA.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2013) Biotechnology, Delft University of 
            Technology, Julianalaan 67, Delft 2628BC, The Netherlands
REFERENCE   3  (bases 1 to 11766)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: IDBA v. 0.20
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11766
                     /organism="Expression vector pUDE215"
                     /mol_type="other DNA"
                     /db_xref="taxon:1431317"
     rep_origin      1..941
                     /label=pMB1 origin
                     /note="pMB1 origin"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(942..1802)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="AHB33290.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(942..1802)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1934..3279
                     /label=2-micron origin
                     /note="2-micron origin"
     rep_origin      1934..3276
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2897..2944)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        3538..3753
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3754..4557
                     /codon_start=1
                     /gene="URA3"
                     /product="orotidine 5-phosphate decarboxylase"
                     /label=URA3
                     /protein_id="AHB33289.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            3754..4557
                     /gene="URA3"
                     /label=URA3
     rep_origin      complement(4688..5143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5284..5300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5310..5328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5353..6357
                     /regulatory_class="promoter"
                     /note="TPI1 promoter"
     CDS             6358..8646
                     /codon_start=1
                     /gene="PFL"
                     /product="pyruvate formate lyase"
                     /label=PFL
                     /protein_id="AHB33291.1"
                     /translation="MTLKASKKAAVPVMQAKTYATAPVITNDAAAKSEIDVEGWIKKHY
                     TPYEGDGSFLAGPTEKTKKLFAKAEEYLAKERANGGLYDVDPHTPSTITSHKPGYLDKE
                     NEVIYGYQTDVPLKRAIKPFGGVNMVKNALKAVNVPMDKEVEHIFSDYRKTHNTAVFDI
                     YSKEMRAGRSNAIMTGLPDGYGRGRIIGDYRRVALYGTDRLIAQKEKDKAELQKKQMDE
                     PTMKLIGEVADQVKALKQLTQMAKSYGIDITKPAKNAREATQFVYFGYLGSIKEQDGAA
                     MSLGRVDAFLDCFFENDLKNGVITESEAQEIIDNLILKLRFARHLRTPEYNDLFAGDPT
                     WVTMSLGGMGSDGRTLVTKTSFRVLNTLYNLGPAPEPNITVLWNKNLPKNFKDFATKVS
                     IDTSSIQYESDALMSARFGDDYGIACCVSAMRIGKDMQFFGARCNLAKLMLYVLNHGKD
                     ERTGKQVGPDFGPVPEGPIPFDWMWETYDKAMDWIAKLYVNTMNVIHFCHDQYCYESLQ
                     MALHDTDVRRLMAFGVAGLSVVADSFSAIKYAKVTPIRDPKTGLTVDFKVEGEFPKFGN
                     DDDRVDFFARTVTDKLINKLRKTPTYRGATHTLSILTITSNVVYGKKTGSTPDGRKAGQ
                     PFAPGCNPMHGREFSGAVASLSSVAKVNYDSCMDGISNTFSIVPNTIGKTLQERQGNLS
                     GLLDGYFSKGAHHLNVNVLKRETLEDAMAHPENYPNLTIRVSGYAVNFVKLTPAQQKEV
                     IARTFHEKM"
     gene            6358..8646
                     /gene="PFL"
                     /label=PFL
     regulatory      8647..9147
                     /regulatory_class="terminator"
                     /note="GND2 terminator"
     regulatory      9152..10152
                     /regulatory_class="promoter"
                     /note="FBA1 promoter"
     CDS             10153..10953
                     /codon_start=1
                     /gene="PFL-AE"
                     /product="pyruvate formate lyase activating enzyme"
                     /label=PFL-AE
                     /protein_id="AHB33292.1"
                     /translation="MPAIVDPTTMDYMEVKGNVHSTESLACLEGPGNRFLLFLNGCAAR
                     CLYCSNPDTWDETVGTPMTVGQLIKKIGNLKNYYINSVGGGGVTVSGGEPLTQFGFLSC
                     FLYAVKKHLNLHTCVETTGQGCTKAWNSVLPHTDLCLVCIKHAIPEKYEQITRTKKLDR
                     CLKFLKELEKRNIPWWCRYVVLPGYTDSKEDIEALIELVKNSPTCERIEFLPYPELGKN
                     KWEELGIEYPLKNIKQLKKSEIKWICDMVREAFKDRNIPVTGDT"
     gene            10153..10953
                     /gene="PFL-AE"
                     /label=PFL-AE
     regulatory      10954..11454
                     /regulatory_class="terminator"
                     /note="PMA1 terminator"
     primer_bind     complement(11455..11471)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(11508..11526)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11547..11563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11571..11587
                     /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)."
     promoter        complement(11595..11625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11640..11661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."