pUDE150 vector (V002347)

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LOCUS       40924_45423        9307 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDE150, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9307)
  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 9307)
  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 9307)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9307)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            (2013) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAY-2013) Biotechnology, Delft University of Technology, 
            Julianalaan 67, Delft 2628BC, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(945..1802)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1803..1907)
                     /label=AmpR promoter
     rep_origin      complement(1934..3276)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3540..3760
                     /label=URA3 promoter
     CDS             3761..4561
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4695..5150)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5295..5311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5321..5339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5358..5605)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5608..5624
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5657..5681
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5697..8306)
                     /codon_start=1
                     /gene="adhE"
                     /product="alcohol-acetaldehyde dehydrogenase"
                     /label=adhE
                     /protein_id="AHB33295.1"
                     /translation="MLTIPEKENRGSKEQEVAIMIDALADKGKKALEALSKKSQEEIDH
                     IVHQMSLAAVDQHMVLAKLAHEETGRGIYEDKAIKNLYASEYIWNSIKDNKTVGIIGED
                     KEKGLTYVAEPIGVICGVTPTTNPTSTTIFKAMIAIKTGNPIIFAFHPSAQESSKRAAE
                     VVLEAAMKAGAPKDIIQWIEVPSIEATKQLMNHKGIALVLATGGSGMVKSAYSTGKPAL
                     GVGPGNVPSYIEKTAHIKRAVNDIIGSKTFDNGMICASEQVVVIDKEIYKDVTNEFKAH
                     QAYFVKKDELQRLENAIMNEQKTGIKPDIVGKSAVEIAELAGIPVPENTKLIIAEISGV
                     GSDYPLSREKLSPVLALVKAQSTKQAFQICEDTLHFGGLGHTAVIHTEDETLQKDFGLR
                     MKACRVLVNTPSAVGGIGDMYNELIPSLTLGCGSYGRNSISHNVSATDLLNIKTIAKRR
                     NNTQIFKVPAQIYFEENAIMSLTTMDKIEKVMIVCDPGMVEFGYTKTVENVLRQRTEQP
                     QIKIFSEVEPNPSTNTVYKGLEMMVDFQPDTIIALGGGSAMDAAKAMWMFFEHPETSFF
                     GAKQKFLDIGKRTYKIGMPENATFICIPTTSGTGSEVTPFAVITDSETNVKYPLADFAL
                     TPDVAIIDPQFVMSVPKSVTADTGMDVLTHAMESYVSVMASDYTRGLSLQAIKLTFEYL
                     KSSVEKGDKVSREKMHNASTLAGMAFANAFLGIAHSIAHKIGGEYGIPHGRANAILLPH
                     IIRYNAKDPQKHALFPKYEFFRADTDYADIAKFLGLKGNTTEALVESLAKAVYELGQSV
                     GIEMNLKSQGVSEEELNESIDRMAELAFEDQCTTANPKEALISEIKDIIQTSYDYKQ"
     gene            complement(5697..8306)
                     /gene="adhE"
                     /label=adhE
     protein_bind    complement(8322..8346)
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(8354..8370)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(8376..9020)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(9049..9067)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9088..9104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(9112..9128)
                     /label=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)."
     promoter        complement(9136..9166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9181..9202)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."