LOCUS       sequence-327682-        8515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of SOD1-AID* (mini auxin inducible 
            degron)-yEGFP and rice auxin receptor gene OsTIR1. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8515)
  AUTHORS   Lu Z, Peng B, Ebert BE, Dumsday G, Vickers CE
  TITLE     Auxin-mediated protein depletion for metabolic engineering in 
            terpene-producing yeast.
  JOURNAL   Nat Commun. 2021 Feb 16;12(1):1051. doi: 10.1038/s41467-021-21313-1.
  PUBMED    33594068
REFERENCE   2  (bases 1 to 8515)
  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"; journalName: "Nat 
            Commun."; date: "2021-02-16"; volume: "12(1):1051. doi"; pages: " 
            10.1038/s41467-021-21313-1"
COMMENT     Sequence Label: pJAID5WY
FEATURES             Location/Qualifiers
     source          1..8515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             946..1077
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     CDS             1087..1797
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1813..2083
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             2579..4306
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     primer_bind     complement(4536..4552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4536..4552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4549..4571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4560..4576
                     /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(join(4584..4590,4591..4608,4609..4614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4767..4784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4938..5526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5018..5037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5697..6488,6489..6557))
                     /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     6320..6339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6558..6662)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6730..6748
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6786..6808)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6908..6927
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7121..7143
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7135..7152
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7136..7152
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8266..8299)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8510..8515,1..408)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
       61 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
      121 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
      181 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
      241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
      301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
      361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      421 actcgagaaa atggttactc aattcaaaac tctaatctaa gttttaatta caaactcgag
      481 aaaatggttc aagcagtcgc agtgttaaag ggtgatgccg gtgtctctgg tgttgtcaag
      541 ttcgaacagg cttccgaatc cgagccaacc actgtctctt acgagatcgc tggtaacagt
      601 cctaacgcag aacgtgggtt ccacattcat gagtttggag atgccaccaa tggttgtgtc
      661 tctgctggtc ctcacttcaa tcctttcaag aagacacatg gtgctccaac tgacgaagtc
      721 agacatgtcg gtgacatggg taacgtaaag acggacgaaa atggtgtggc caagggctcc
      781 ttcaaggact ctttgatcaa gcttatcggt cctacctccg ttgtaggcag aagcgtcgtt
      841 atccacgccg gccaagatga cttaggtaag ggtgacactg aagaatcttt gaagactggt
      901 aatgccggtc caagaccagc ctgtggtgtc attggtctaa ccaacccaaa agacccagct
      961 aaacctccag ccaaggccca agtcgtcggc tggccaccag ttaggtctta cagaaagaac
     1021 gtaatggtta gttgtcaaaa gtcctccggt ggtccagaag ctgctgcctt tgttaagatg
     1081 ggatcctcta aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat
     1141 ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac
     1201 ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc
     1261 ttagtcacta ctttcggtta tggtgttcaa tgttttgcga gatacccaga tcatatgaaa
     1321 caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt
     1381 ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta
     1441 gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac
     1501 aaattggaat acaactataa ctctcacaat gtttacatca tggctgacaa acaaaagaat
     1561 ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt tcaattagct
     1621 gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat
     1681 tacttatcca ctcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc
     1741 ttgttagaat ttgttactgc tgctggtatt acccatggta tggatgaatt gtacaaaaga
     1801 tcttaaacta gtattgaatt gaattgaaat cgatagatca atttttttct tttctctttc
     1861 cccatccttt acgctaaaat aatagtttat tttatttttt gaatattttt tatttatata
     1921 cgtatatata gactattatt tatcttttaa tgattattaa gatttttatt aaaaaaaaat
     1981 tcgctcctct tttaatgcct ttatgcagtt tttttttccc attcgatatt tctatgttcg
     2041 ggttcagcgt attttaagtt taataactcg aaaattctgc gttcgttaaa gctttcgaga
     2101 aggatattat ttgcggccgc gattgcaaga attgatctcc ctcatcgatg tcataaatca
     2161 gtaaatataa acattaaata cattacacgt agccacccgc tgtatagcat aaaactaact
     2221 taaaattatt catatgcgtt tcccggggcc gaagcgttat tgccgatatt cggtagccga
     2281 ttccgctggc tttttttttt tgaacgaggc aaaaaagttt cgtgactgaa cttcttcgtc
     2341 actggttttg ttgataattt ttatcgtttt ttacttcgag attgaaaaca gtgttctgaa
     2401 ttttagcaac gatatcgcac cgcacgttta gcctgatcaa accttatata tacctctacc
     2461 ttaaaatata tatatatata gtattcgaat tccgaattgt gtgtctccgc ggttagtgat
     2521 tgttatacac aaacagaata caggaaagta aatcaataca ataataaatt ctagcaaaat
     2581 gacctatttc ccagaagaag tcgttgaaca catcttctct ttcttgccag ctcaaagaga
     2641 tagaaatact gtttccttgg tttgtaaggt ttggtatgaa atcgaaagat tgagtagacg
     2701 tggtgttttc gtcggtaatt gttacgctgt tagagctggt agagttgctg ctagattccc
     2761 aaatgttaga gccttgactg tcaaaggtaa accacatttt gctgacttta acttggtccc
     2821 accagattgg ggtggttatg ctggtccatg gattgaagct gccgctagag gttgtcacgg
     2881 tttggaagaa ttgcgtatga aaagaatggt cgtttctgac gaatctttgg aattattggc
     2941 ccgttctttt ccaagattca gagctttggt tttgatctct tgtgaaggtt tcagtactga
     3001 cggtttggct gccgttgctt cccattgtaa attattgaga gaattggact tgcaagaaaa
     3061 tgaagtcgaa gatagaggtc caagatggtt atcttgtttt ccagactctt gtacttcttt
     3121 ggtctctttg aacttcgctt gtattaaggg tgaagtcaac gctggttctt tagaaagatt
     3181 ggtctcccgt tctccaaact tacgttcttt gagattgaat cgttctgttt ccgttgatac
     3241 cttggctaaa atcttattga gaaccccaaa cttggaagac ttaggtactg gtaacttgac
     3301 cgacgatttc caaactgaat cctactttaa attaacttct gctttagaaa agtgtaagat
     3361 gttgagatcc ttatctggtt tttgggacgc ctctccagtc tgtttatctt ttatctatcc
     3421 attgtgtgct caattaactg gtttaaactt atcttacgct ccaaccttgg atgctagtga
     3481 cttaaccaag atgattagta gatgtgttaa gttacaaaga ttatgggttt tagattgtat
     3541 ttccgataag ggtttacaag tcgttgctag tagctgtaag gatttgcaag aattaagagt
     3601 ctttccaagt gatttttacg ttgctggtta cagcgctgtt actgaagaag gtttggtcgc
     3661 tgtttctttg ggttgtccaa agttgaacag cttattgtac ttttgtcatc aaatgactaa
     3721 tgctgcctta gtcactgttg ctaagaactg tccaaacttc acccgtttca gattgtgtat
     3781 tttggaacca ggtaaaccag acgttgtcac ctcccaacca ttggatgaag gtttcggtgc
     3841 tatcgttaga gaatgtaagg gtttgcaaag attatctatc tctggtttgt taactgacaa
     3901 agttttcatg tacatcggta aatatgccaa acaattagaa atgttgagca ttgctttcgc
     3961 tggtgactct gacaagggta tgatgcacgt tatgaatggt tgtaagaatt tgagaaagtt
     4021 ggaaattaga gattctccat tcggtgacgc tgccttattg ggtaattttg ctagatacga
     4081 aactatgaga agtttgtgga tgagttcctg taacgttacc ttgaaaggtt gtcaagtctt
     4141 agccagcaag atgccaatgt tgaacgtcga agtcatcaac gaaagagacg gtagtaacga
     4201 aatggaagaa aatcacggtg acttaccaaa ggtagaaaag ttgtacgtat acagaactac
     4261 tgctggtgct agagatgatg ctccaaattt cgtaaagatt ttgtaaagga ctatttgcaa
     4321 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     4381 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     4441 actcacaaat tagagcttca atttaattat atcagttatt acccgggaat ctcggtcgta
     4501 atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4561 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4621 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4741 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4801 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5341 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5821 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6661 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6721 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6781 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6841 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6901 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     6961 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     7021 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7081 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7141 acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa cagaaggaag
     7201 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     7261 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa
     7321 gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca agtccggttg
     7381 catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct tctcttgacg
     7441 ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac atttgccttt
     7501 tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc gttccattga
     7561 aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc gccgatatcg
     7621 gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa tggtctgata
     7681 tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa caaggtgcgc
     7741 aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct tccaagggtt
     7801 ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt gataaagatt
     7861 tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg tttgattggc
     7921 taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt cagcagtaca
     7981 gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt
     8041 tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct ggatgggaag
     8101 cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat agaacaaatc
     8161 acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc aacttcatct
     8221 tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac ttcgtatagc
     8281 atacattata cgaagttata ttaagggttg ctcgaccaga aagcgaccac ccaactttgg
     8341 ctgataatag cgtataaaca atgcatactt tgtacgttca aaatacaatg cagtagatat
     8401 atttatgcat attacatata atacatatca cataggaagc aacaggcgcg ttggactttt
     8461 aattttcgag gaccgcgaat ccttacatca cacccaatcc cccacaagtg atccc
//
