LOCUS       sequence-327152-        7865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of yEGFP and rice auxin receptor gene OsTIR1.
            OsTIR1 is codon-optimised for expression in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7865)
  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 7865)
  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: pJAID9L
FEATURES             Location/Qualifiers
     source          1..7865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             431..1141
                     /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      1157..1427
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             1923..3650
                     /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(3886..3902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3886..3902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3899..3921)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3910..3926
                     /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(3934..3940,3941..3958,3959..3964))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3979..4000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4117..4134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4288..4876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4368..4387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5047..5838,5839..5907))
                     /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     5670..5689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5908..6012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6080..6098
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6136..6158)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6258..6277
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6471..6493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6485..6502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6486..6502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7616..7649)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(7860..7865,1..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 aatgggatcc tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt
      481 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac
      541 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc
      601 aaccttagtc actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat
      661 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat
      721 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac
      781 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg
      841 tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa
      901 gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt
      961 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa
     1021 ccattactta tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat
     1081 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa
     1141 aagatcttaa actagtattg aattgaattg aaatcgatag atcaattttt ttcttttctc
     1201 tttccccatc ctttacgcta aaataatagt ttattttatt ttttgaatat tttttattta
     1261 tatacgtata tatagactat tatttatctt ttaatgatta ttaagatttt tattaaaaaa
     1321 aaattcgctc ctcttttaat gcctttatgc agtttttttt tcccattcga tatttctatg
     1381 ttcgggttca gcgtatttta agtttaataa ctcgaaaatt ctgcgttcgt taaagctttc
     1441 gagaaggata ttatttgcgg ccgcgattgc aagaattgat ctccctcatc gatgtcataa
     1501 atcagtaaat ataaacatta aatacattac acgtagccac ccgctgtata gcataaaact
     1561 aacttaaaat tattcatatg cgtttcccgg ggccgaagcg ttattgccga tattcggtag
     1621 ccgattccgc tggctttttt tttttgaacg aggcaaaaaa gtttcgtgac tgaacttctt
     1681 cgtcactggt tttgttgata atttttatcg ttttttactt cgagattgaa aacagtgttc
     1741 tgaattttag caacgatatc gcaccgcacg tttagcctga tcaaacctta tatatacctc
     1801 taccttaaaa tatatatata tatagtattc gaattccgaa ttgtgtgtct ccgcggttag
     1861 tgattgttat acacaaacag aatacaggaa agtaaatcaa tacaataata aaagctagca
     1921 aaatgaccta tttcccagaa gaagtcgttg aacacatctt ctctttcttg ccagctcaaa
     1981 gagatagaaa tactgtttcc ttggtttgta aggtttggta tgaaatcgaa agattgagta
     2041 gacgtggtgt tttcgtcggt aattgttacg ctgttagagc tggtagagtt gctgctagat
     2101 tcccaaatgt tagagccttg actgtcaaag gtaaaccaca ttttgctgac tttaacttgg
     2161 tcccaccaga ttggggtggt tatgctggtc catggattga agctgccgct agaggttgtc
     2221 acggtttgga agaattgcgt atgaaaagaa tggtcgtttc tgacgaatct ttggaattat
     2281 tggcccgttc ttttccaaga ttcagagctt tggttttgat ctcttgtgaa ggtttcagta
     2341 ctgacggttt ggctgccgtt gcttcccatt gtaaattatt gagagaattg gacttgcaag
     2401 aaaatgaagt cgaagataga ggtccaagat ggttatcttg ttttccagac tcttgtactt
     2461 ctttggtctc tttgaacttc gcttgtatta agggtgaagt caacgctggt tctttagaaa
     2521 gattggtctc ccgttctcca aacttacgtt ctttgagatt gaatcgttct gtttccgttg
     2581 ataccttggc taaaatctta ttgagaaccc caaacttgga agacttaggt actggtaact
     2641 tgaccgacga tttccaaact gaatcctact ttaaattaac ttctgcttta gaaaagtgta
     2701 agatgttgag atccttatct ggtttttggg acgcctctcc agtctgttta tcttttatct
     2761 atccattgtg tgctcaatta actggtttaa acttatctta cgctccaacc ttggatgcta
     2821 gtgacttaac caagatgatt agtagatgtg ttaagttaca aagattatgg gttttagatt
     2881 gtatttccga taagggttta caagtcgttg ctagtagctg taaggatttg caagaattaa
     2941 gagtctttcc aagtgatttt tacgttgctg gttacagcgc tgttactgaa gaaggtttgg
     3001 tcgctgtttc tttgggttgt ccaaagttga acagcttatt gtacttttgt catcaaatga
     3061 ctaatgctgc cttagtcact gttgctaaga actgtccaaa cttcacccgt ttcagattgt
     3121 gtattttgga accaggtaaa ccagacgttg tcacctccca accattggat gaaggtttcg
     3181 gtgctatcgt tagagaatgt aagggtttgc aaagattatc tatctctggt ttgttaactg
     3241 acaaagtttt catgtacatc ggtaaatatg ccaaacaatt agaaatgttg agcattgctt
     3301 tcgctggtga ctctgacaag ggtatgatgc acgttatgaa tggttgtaag aatttgagaa
     3361 agttggaaat tagagattct ccattcggtg acgctgcctt attgggtaat tttgctagat
     3421 acgaaactat gagaagtttg tggatgagtt cctgtaacgt taccttgaaa ggttgtcaag
     3481 tcttagccag caagatgcca atgttgaacg tcgaagtcat caacgaaaga gacggtagta
     3541 acgaaatgga agaaaatcac ggtgacttac caaaggtaga aaagttgtac gtatacagaa
     3601 ctactgctgg tgctagagat gatgctccaa atttcgtaaa gattttgtaa ctcgagagga
     3661 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     3721 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     3781 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat
     3841 ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6001 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6061 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6241 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6481 acgttgtaaa acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa
     6541 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     6601 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc
     6661 aggaaacgaa gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca
     6721 agtccggttg catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct
     6781 tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac
     6841 atttgccttt tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc
     6901 gttccattga aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc
     6961 gccgatatcg gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa
     7021 tggtctgata tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa
     7081 caaggtgcgc aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct
     7141 tccaagggtt ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt
     7201 gataaagatt tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg
     7261 tttgattggc taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt
     7321 cagcagtaca gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc
     7381 agaggacttt tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct
     7441 ggatgggaag cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat
     7501 agaacaaatc acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc
     7561 aacttcatct tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac
     7621 ttcgtatagc atacattata cgaagttata ttaagggttg ctcgaccaga aagcgaccac
     7681 ccaactttgg ctgataatag cgtataaaca atgcatactt tgtacgttca aaatacaatg
     7741 cagtagatat atttatgcat attacatata atacatatca cataggaagc aacaggcgcg
     7801 ttggactttt aattttcgag gaccgcgaat ccttacatca cacccaatcc cccacaagtg
     7861 atccc
//
