LOCUS       sequence-370810-        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knocking out ALD6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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: pIALD2E
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(64..652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(144..163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(823..1614,1615..1683))
                     /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     1446..1465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1684..1788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1815..3157
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2778..2825)
                     /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     3198..3216
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3254..3276)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3376..3395
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3416..3697
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3698..4372
                     /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"
ORIGIN
        1 tttaaattga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
       61 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1801 aaaagtgcca cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     1861 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     1921 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     1981 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2041 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
     2101 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
     2161 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
     2221 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
     2281 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
     2341 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
     2401 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     2461 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     2521 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     2581 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     2641 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     2701 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     2761 gttttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
     2821 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     2881 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     2941 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     3001 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     3061 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     3121 catccttcaa tgctatcatt tcctttgata ttggatcata ctaagaaacc attattatca
     3181 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     3241 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     3301 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3361 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat agatcaacga
     3421 cattactata tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac
     3481 tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg
     3541 tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc
     3601 ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg
     3661 agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg
     3721 tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc
     3781 tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat
     3841 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     3901 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     3961 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata
     4021 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     4081 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc
     4141 aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga
     4201 gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga
     4261 tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg
     4321 tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac
     4381 tgagtagtat ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc
     4441 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     4501 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4561 aaaatccctt aatttaaatg taataagaag tttggtaata ttcaattcga agtgttcagt
     4621 cttttacttc tcttgtttta tagaagaaaa aacatcaaga aacatcttta acatacacaa
     4681 acacatacta tcagaataca atgactaagc tacactttga cactgctgaa ccagtcaaga
     4741 gtgcagtgta ccaacctgca tttctttccg tcatatacac aaaatacttt catataaact
     4801 tacttggtct tacgtcataa ataaatatgt atacatataa attaaaaaat ttggttttat
     4861 atttttacaa aaagaatcgt ttacttcatt tctccctttt aagcgataca atccatgaaa
     4921 aaagagaaaa agagagaaca ggcttgtgcc ttctttaaaa catcccacac aaaatcatat
     4981 tgaattgaa
//
