LOCUS       40924_45433        8107 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDE152, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8107)
  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 8107)
  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 8107)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8107)
  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..8107
                     /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..7106)
                     /codon_start=1
                     /gene="lin1129"
                     /product="lin1129"
                     /label=lin1129
                     /protein_id="AHB33301.1"
                     /translation="MESLELEQLVKKVLLEKLAEQKEVPTKTTTQGAKSGVFDTVDEAV
                     QAAVIAQNCYKEKSLEERRNVVKAIREALYPEIETIATRAVAETGMGNVTDKILKNTLA
                     IEKTPGVEDLYTEVATGDNGMTLYELSPYGVIGAVAPSTNPTETLICNSIGMLAAGNAV
                     FYSPHPGAKNISLWLIEKLNTIVRDSCGIDNLIVTVAKPSIQAAQEMMNHPKVPLLVIT
                     GGPGVVLQAMQSGKKVIGAGAGNPPSIVDETANIEKAAADIVDGASFDHNILCIAEKSV
                     VAVDSIADFLLFQMEKNGALHVTNPSDIQKLEKVAVTDKGVTNKKLVGKSATEILKEAG
                     IACDFTPRLIIVETEKSHPFATVELLMPIVPVVRVPDFDEALEVAIELEQGLHHTATMH
                     SQNISRLNKAARDMQTSIFVKNGPSFAGLGFRGEGSTTFTIATPTGEGTTTARHFARRR
                     RCVLTDGFSIR"
     gene            complement(5697..7106)
                     /gene="lin1129"
                     /label=lin1129
     protein_bind    complement(7122..7146)
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7154..7170)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7176..7820)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        complement(7849..7867)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7888..7904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7912..7928)
                     /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(7936..7966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7981..8002)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     1981 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2041 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2101 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2161 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2221 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2281 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     2341 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     2401 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     2461 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     2521 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     2581 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     2641 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     2701 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     2761 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     2821 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     2881 ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     2941 cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca
     3001 gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa
     3061 gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga
     3121 tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct
     3181 tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc
     3241 atccttcaat gctatcattt cctttgatat tggatcatac taagaaacca ttattatcat
     3301 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     3361 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3421 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3481 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata ccacagcttt
     3541 tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc tttgaaattt
     3601 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     3661 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     3721 actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     3781 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     3841 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     3901 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     3961 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     4021 actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     4081 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     4141 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     4201 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     4261 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     4321 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     4381 tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg
     4441 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     4501 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     4561 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     4621 ttaattatat cagttattac cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4681 aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     4741 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4801 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4861 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4921 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4981 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5041 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5101 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcgccattc
     5161 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5221 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5281 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     5341 cgaattgggt accggccgca aattaaagcc ttcgagcgtc ccaaaacctt ctcaagcaag
     5401 gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga
     5461 aatataaata acgttcttaa tactaacata actataaaaa aataaatagg gacctagact
     5521 tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaatgt
     5581 aagcgtgaca taactaatta catgactcga ggtcgacggt atcgataagc ttgatatcga
     5641 attcctgcag cccatcacca ctttgtacaa gaaagctggg tgtcgactag gaattcttat
     5701 ctgatagaga aaccgtcagt caaaacacat cttcttcttc tagcgaagtg tctagcagta
     5761 gtagtacctt caccagttgg agtagcgata gtgaaagtag tagaaccttc acctctgaaa
     5821 cccaaaccag cgaaagatgg accgttctta acgaagatag aagtttgcat gtctctagca
     5881 gccttgttca atctagagat gttttgagag tgcatagtag cagtgtggtg caaaccttgt
     5941 tccaattcga tagcaacttc caaagcttcg tcgaagtctg gaactctaac aactggaacg
     6001 attggcatca acaattcaac agtagcgaat gggtgagact tttcagtttc aacgatgatc
     6061 aatcttggag tgaagtcaca agcgatacca gcttccttca agatttcagt agcagactta
     6121 ccaaccaact tcttgttagt aacacccttg tcagtaacag caaccttttc caacttttgg
     6181 atgtcagatg ggttagtaac gtgcaaagca ccgttctttt ccatttggaa caacaagaag
     6241 tcagcgatag agtcaacagc aacaacagac ttttcagcga tacacaagat gttgtggtcg
     6301 aaagaagcac cgtcaacgat gtcagcagca gccttttcga tgttagcagt ttcgtcaacg
     6361 atagatggtg ggttaccagc accagcaccg ataaccttct taccagattg catagcttgc
     6421 aaaacaacac ctggaccacc agtgataacc aacaatggaa cctttgggtg gttcatcatt
     6481 tcttgagcag cttggataga tggcttagca acagtaacga tcaagttgtc gataccacaa
     6541 gagtctctaa cgatagtgtt caacttttcg atcaaccaca aagagatgtt cttagcacct
     6601 gggtgtggag agtagaaaac agcgttacca gcagccaaca taccgataga gttacagatc
     6661 aaagtttcag ttgggttagt agatggagca acagcaccga taacaccgta tggagacaat
     6721 tcgtacaaag tcataccgtt gtcaccagta gcaacttcag tgtacaagtc ttcaacacct
     6781 ggagtctttt cgatagccaa agtgttcttc aagatcttgt cagtaacgtt acccatacca
     6841 gtttcagcaa cagctctagt agcgatagtt tcgatttctg ggtacaaagc ttctctgata
     6901 gccttaacaa cgtttcttct ttcttccaaa gacttttcct tgtaacagtt ttgagcgata
     6961 acagcagctt gaacagcttc gtcaacagtg tcgaaaacac cagacttagc accttgagta
     7021 gtagtcttag ttggaacttc cttttgttca gccaactttt ccaacaaaac cttcttaacc
     7081 aattgttcca attccaaaga ttccatggat cctatctgca gagcctgctt ttttgtacaa
     7141 acttgtgatg ggggatccac tagttctaga atccgtcgaa actaagttct ggtgttttaa
     7201 aactaaaaaa aagactaact ataaaagtag aatttaagaa gtttaagaaa tagatttaca
     7261 gaattacaat caatacctac cgtctttata tacttattag tcaagtaggg gaataatttc
     7321 agggaactgg tttcaacctt ttttttcagc tttttccaaa tcagagagag cagaaggtaa
     7381 tagaaggtgt aagaaaatga gatagataca tgcgtgggtc aattgccttg tgtcatcatt
     7441 tactccaggc aggttgcatc actccattga ggttgtgccc gttttttgcc tgtttgtgcc
     7501 cctgttctct gtagttgcgc taagagaatg gacctatgaa ctgatggttg gtgaagaaaa
     7561 caatattttg gtgctgggat tctttttttt tctggatgcc agcttaaaaa gcgggctcca
     7621 ttatatttag tggatgccag gaataaacct gttcacccag gcaccttcgg tgttatatat
     7681 tctgtgtaac ccgcccccta ttttgggcat gtacgggtta cagcagaatt aaaaggctaa
     7741 ttttttgact aaataaagtt aggaaaatca ctactattaa ttatttacgt attctttgaa
     7801 atggcgagta ttgataatga taaactcgaa gagctccagc ttttgttccc tttagtgagg
     7861 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7921 gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta
     7981 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     8041 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     8101 tgggcgc
//
