LOCUS       sequence-370809-        7487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knocking out ALD6 and expressing anacetylating aldehyde 
            dehydrogenase (Lactobacillus reuteri pduP ) in Saccharomyces 
            cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7487)
  AUTHORS   Hayat IF, Plan M, Ebert BE, Dumsday G, Vickers CE, Peng B
  TITLE     Auxin-mediated induction of GAL promoters by conditional degradation
            of Mig1p improves sesquiterpene production in Saccharomyces 
            cerevisiae with engineered acetyl-CoA synthesis.
  JOURNAL   Microb Biotechnol. 2021 Nov;14(6):2627-2642. doi: 
            10.1111/1751-7915.13880. Epub 2021 Sep 9.
  PUBMED    34499421
REFERENCE   2  (bases 1 to 7487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/1751-7915.13880"; journalName: "Microb Biotechnol"; date: 
            "2021-11"; volume: "14"; issue: "6"; pages: "2627-2642"
COMMENT     Sequence Label: pIALD2S
FEATURES             Location/Qualifiers
     source          1..7487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(299..887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(379..398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1058..1849,1850..1918))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1681..1700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1919..2023)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2050..3392
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3013..3060)
                     /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)."
     primer_bind     3433..3451
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3489..3511)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3611..3630
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3651..3932
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3933..4607
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        5024..5728
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 cctgcatttc tttccgtcat atacacaaaa tactttcata taaacttact tggtcttacg
       61 tcataaataa atatgtatac atataaatta aaaaatttgg ttttatattt ttacaaaaag
      121 aatcgtttac ttcatttctc ccttttaagc gatacaatcc atgaaaaaag agaaaaagag
      181 agaacaggct tgtgccttct ttaaaacatc ccacacaaaa tcatattgaa ttgaatttaa
      241 attgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      721 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     1981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2041 tgccacctga acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2101 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2161 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2221 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2281 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2341 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     2401 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     2461 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     2521 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     2581 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     2641 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     2701 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     2761 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     2821 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     2881 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     2941 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt
     3001 caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc
     3061 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     3121 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     3181 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     3241 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     3301 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     3361 ttcaatgcta tcatttcctt tgatattgga tcatactaag aaaccattat tatcatgaca
     3421 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3481 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3541 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3601 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagatc aacgacatta
     3661 ctatatatat aatataggaa gcatttaata gaacagcatc gtaatatatg tgtactttgc
     3721 agttatgacg ccagatggca gtagtggaag atattcttta ttgaaaaata gcttgtcacc
     3781 ttacgtacaa tcttgatccg gagcttttct ttttttgccg attaagaatt aattcggtcg
     3841 aaaaaagaaa aggagagggc caagagggag ggcattggtg actattgagc acgtgagtat
     3901 acgtgattaa gcacacaaag gcagcttgga gtatgtctgt tattaatttc acaggtagtt
     3961 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag
     4021 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
     4081 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
     4141 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
     4201 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
     4261 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
     4321 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
     4381 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
     4441 ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
     4501 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
     4561 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaataggtt attactgagt
     4621 agtatttatt taagtattgt ttgtgcactt gccgatctat gcggtgtgaa ataccgcaca
     4681 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     4741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4801 cccttaattt aaatgtaata agaagtttgg taatattcaa ttcgaagtgt tcagtctttt
     4861 acttctcttg ttttatagaa gaaaaaacat caagaaacat ctttaacata cacaaacaca
     4921 tactatcaga atacaatgac taagctacac tttgacactg ctgaaccagt caagaggatc
     4981 cgctcttttc cgattttttt ctaaaccgtg gaatatttcg gatatccttt tgttgtttcc
     5041 gggtgtacaa tatggacttc ctcttttctg gcaaccaaac ccatacatcg ggattcctat
     5101 aataccttcg ttggtctccc taacatgtag gtggcggagg ggagatatac aatagaacag
     5161 ataccagaca agacataatg ggctaaacaa gactacacca attacactgc ctcattgatg
     5221 gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc atatcgaagt
     5281 ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat gcaacttctt
     5341 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
     5401 aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt
     5461 tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattca
     5521 cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga
     5581 aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt
     5641 ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt
     5701 tcaagctata ccaagcatac aatcaactat ctcatataca ggtaccaaaa tgcagattaa
     5761 cgatattgaa agtgctgtac gcaaaattct tgcagaagaa ttagataatg ctagctctaa
     5821 cagtgcggca gttactaatg atgatggaca tcgtggtatt tttactaatg ttgatgacgc
     5881 aattgctgct gctaaagctg cccaagaagt atatcgtgac aagccaattg cagtgcggca
     5941 acaggtgatt gatgctatca aggaaggttt ccgtccttac attgaaaaga tggcaaagga
     6001 cattaaagaa gaaaccggaa tgggaacagt cgaagcaaag attgctaagt tgcacaatgc
     6061 tttgtacaac actcctggtc cagaaattct tgaaccggtt gtcgaaaatg gtgatggcgg
     6121 aatggttatg tatgaacggt tgccatacgg tgttatcggt gctgttggtc caagtactaa
     6181 cccatccgaa acagttattg ccaatgcaat tatgatgctt gcaggtggaa atacccttta
     6241 ctttggtgct caccctggtg ctaagaatat cactcgttgg acgattgaaa agatgaatga
     6301 tttcattgct gatgcaactg gtcttcacaa tttagttgta agcattgaaa caccaacaat
     6361 tgaatctgtt caacaaatga tgcaacatcc tgatattgct atgcttgccg taaccggtgg
     6421 accagctgtt gttcaccaag caatgactag cggtaagaag gctgttggtg ctggccctgg
     6481 taatcctcct gcaatggttg atgctaccgc tgatattgac ttagctgctc ataatattat
     6541 tgcttcagct tcatttgata atgacatttt atgtactgct gaaaaggaag ttgttgccga
     6601 aagtagtatt aaggatgaat taatccgcaa gatgcaagat gaaggtgcat ttgttgttaa
     6661 ccgtgaacag gctgacaaac tcgccgatat gtgcatccaa gacaacggta ctcctgatcg
     6721 gaagtttgtt ggaaaggatg caacctacat cttagatcaa gctaatattc catatactgg
     6781 tcacccggta gaaattattt gcgaattgcc aaaggaacac ccattggtaa tgactgaaat
     6841 gttgatgcca atcttaccag ttgtttcttg cccaacattt gatgatgttt tgaagactgc
     6901 tgttgaagta gaacaaggaa accaccatac tgcaactatt cactccaaca acttaaagca
     6961 tattaataat gctgctcacc ggatgcaatg ttcgatcttc gttgttaacg gtccatccta
     7021 tgttggaaca ggtgttgctg ataatggtgc tcactcaggt gcttcagcat taacaattgc
     7081 tacgccaact ggtgaaggaa cttgtactgc aagaaccttt acgcgtcgtg ttcgtttgaa
     7141 ttcgccacaa ggattctcag ttcgtaattg gtactaagcg atttaatctc taattattag
     7201 ttaaagtttt ataagcattt ttatgtaacg aaaaataaat tggttcatat tattactgca
     7261 ctgtcactta ccatggaaag accagacaag aagttgccga cacgacagtc tgttgaattg
     7321 gcttaagtct gggtccgctt ctttacaaat ttgaagaatt tctcttaaac gatatgtata
     7381 ttcttttcgt tggaaaagat gtcttccaaa aaaaaaaacc gatgaattag tggaaccaag
     7441 gaaaaaaaaa gaggtatcct tgattaagga acagcgcagt gtaccaa
//
