LOCUS       Exported               12576 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDE065, complete sequence.
ACCESSION   JX268039
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12576)
  AUTHORS   Koopman F, Beekwilder J, Crimi B, Houwelingen A, Hall RD, Bosch D, 
            van Maris AJ, Pronk JT, Daran JM.
  TITLE     De novo production of the flavonoid naringenin in engineered 
            Saccharomyces cerevisiae
  JOURNAL   Microb. Cell Fact. 11 (1), 155 (2012)
  PUBMED    23216753
REFERENCE   2  (bases 1 to 12576)
  AUTHORS   Koopman F, Beekwilder J, Crimi B, van Houwelingen A, Hall RD, Bosch 
            D, Maris AJA., Pronk JT, Daran J-M.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUL-2012) Platform Green Synthetic Biolog, Delft 2600,
            The Netherlands
REFERENCE   3  (bases 1 to 12576)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Name: pUDI065.
FEATURES             Location/Qualifiers
     source          1..12576
                     /organism="Expression vector pUDE065"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:1269199"
     CDS             complement(5..1111)
                     /codon_start=1
                     /product="auxotrophic marker"
                     /label=auxotrophic marker
                     /note="LEU2"
                     /protein_id="AGA15817.1"
                     /translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
                     ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKEL
                     QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
                     EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
                     NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
                     IRTGDLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(5..1099)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1112..1516)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     2043..2059
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2069..2087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2102..2299
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      complement(2102..2296)
                     /regulatory_class="terminator"
                     /operon="TEF1 terminator"
     CDS             complement(2358..4043)
                     /codon_start=1
                     /product="4-CL3"
                     /label=4-CL3
                     /protein_id="AGA15814.1"
                     /translation="MITAALHEPQIHKPTDTSVVSDDVLPHSPPTPRIFRSKLPDIDIP
                     NHLPLHTYCFEKLSSVSDKPCLIVGSTGKSYTYGETHLICRRVASGLYKLGIRKGDVIM
                     ILLQNSAEFVFSFMGASMIGAVSTTANPFYTSQELYKQLKSSGAKLIITHSQYVDKLKN
                     LGENLTLITTDEPTPENCLPFSTLITDDETNPFQETVDIGGDDAAALPFSSGTTGLPKG
                     VVLTHKSLITSVAQQVDGDNPNLYLKSNDVILCVLPLFHIYSLNSVLLNSLRSGATVLL
                     MHKFEIGALLDLIQRHRVTIAALVPPLVIALAKNPTVNSYDLSSVRFVLSGAAPLGKEL
                     QDSLRRRLPQAILGQGYGMTEAGPVLSMSLGFAKEPIPTKSGSCGTVVRNAELKVVHLE
                     TRLSLGYNQPGEICIRGQQIMKEYLNDPEATSATIDEEGWLHTGDIGYVDEDDEIFIVD
                     RLKEVIKFKGFQVPPAELESLLINHHSIADAAVVPQNDEVAGEVPVAFVVRSNGNDITE
                     EDVKEYVAKQVVFYKRLHKVFFVASIPKSPSGKILRKDLKAKLC"
     regulatory      complement(4055..4458)
                     /regulatory_class="promoter"
                     /note="TEF1 promoter"
     promoter        complement(4059..4458)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     4735..4751
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4758..4776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4799..5706
                     /regulatory_class="promoter"
                     /note="TPI1 promoter"
     CDS             5713..6900
                     /codon_start=1
                     /product="CHS3"
                     /label=CHS3
                     /protein_id="AGA15815.1"
                     /translation="MVMAGASSLDEIRQAQRADGPAGILAIGTANPENHVLQAEYPDYY
                     FRITNSEHMTDLKEKFKRMCDKSTIRKRHMHLTEEFLKENPHMCAYMAPSLDTRQDIVV
                     VEVPKLGKEAAVKAIKEWGQPKSKITHVVFCTTSGVDMPGADYQLTKLLGLRPSVKRLM
                     MYQQGCFAGGTVLRIAKDLAENNRGARVLVVCSEITAVTFRGPSDTHLDSLVGQALFSD
                     GAAALIVGSDPDTSVGEKPIFEMVSAAQTILPDSDGAIDGHLREVGLTFHLLKDVPGLI
                     SKNIVKSLDEAFKPLGISDWNSLFWIAHPGGPAILDQVEIKLGLKEEKMRATRHVLSEY
                     GNMSSACVLFILDEMRRKSAKDGVATTGEGLEWGVLFGFGPGLTVETVVLHSVPL"
     regulatory      6907..7230
                     /regulatory_class="terminator"
                     /operon="ADH1 terminator"
     terminator      6907..7094
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        7247..7891
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     regulatory      7364..7903
                     /regulatory_class="promoter"
                     /note="TDH3 promoter"
     CDS             7909..8649
                     /codon_start=1
                     /product="CHI1"
                     /label=CHI1
                     /protein_id="AGA15816.1"
                     /translation="MSSSNACASPSPFPAVTKLHVDSVTFVPSVKSPASSNPLFLGGAG
                     VRGLDIQGKFVIFTVIGVYLEGNAVPSLSVKWKGKTTEELTESIPFFREIVTGAFEKFI
                     KVTMKLPLTGQQYSEKVTENCVAIWKQLGLYTDCEAKAVEKFLEIFKEETFPPGSSILF
                     ALSPTGSLTVAFSKDDSIPETGIAVIENKLLAEAVLESIIGKNGVSPGTRLSVAERLSQ
                     LMMKNKDEKEVSDHSVEEKLAKEN"
     terminator      8662..8909
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     regulatory      8669..8908
                     /regulatory_class="terminator"
                     /operon="CYC1 terminator"
     promoter        complement(8946..8964)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8985..9001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9009..9025
                     /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(9033..9063)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      9372..9991
                     /label=pBR33 Coli replication origin
                     /note="pBR33 Coli replication origin"
     rep_origin      complement(9387..9975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10146..11006)
                     /codon_start=1
                     /product="ampicilin resistance"
                     /label=ampicilin resistance
                     /note="bla"
                     /protein_id="AGA15818.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(11007..11111)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    11253..11964
                     /note="partial PYK2 gene; for integration into the PYK2 
                     locus"