LOCUS       sequence-327153-        8180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of yEGFP-AID*-CUP1 and rice auxin receptor 
            gene OsTIR1. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8180)
  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 8180)
  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: pJAID9L104
FEATURES             Location/Qualifiers
     source          1..8180
                     /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"
     CDS             1148..1279
                     /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"
     terminator      1472..1742
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             2238..3965
                     /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(4201..4217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4201..4217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4214..4236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4225..4241
                     /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(4249..4255,4256..4273,4274..4279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4294..4315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4432..4449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5362..6153,6154..6222))
                     /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     5985..6004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6223..6327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6395..6413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6451..6473)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6573..6592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6786..6808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6800..6817
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6801..6817
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7931..7964)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8175..8180,1..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 aagatctcca aaagacccag ctaaacctcc agccaaggcc caagtcgtcg gctggccacc
     1201 agttaggtct tacagaaaga acgtaatggt tagttgtcaa aagtcctccg gtggtccaga
     1261 agctgctgcc tttgttaaga tgttcagcga attaattaac ttccaaaatg aaggtcatga
     1321 gtgccaatgc caatgtggta gctgcaaaaa taatgaacaa tgccaaaaat catgtagctg
     1381 cccaacgggg tgtaacagcg acgacaaatg cccctgcggt aacaagtctg aagaaaccaa
     1441 gaagtcatgc tgctctggga aataaactag tattgaattg aattgaaatc gatagatcaa
     1501 tttttttctt ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg
     1561 aatatttttt atttatatac gtatatatag actattattt atcttttaat gattattaag
     1621 atttttatta aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca
     1681 ttcgatattt ctatgttcgg gttcagcgta ttttaagttt aataactcga aaattctgcg
     1741 ttcgttaaag ctttcgagaa ggatattatt tgcggccgcg attgcaagaa ttgatctccc
     1801 tcatcgatgt cataaatcag taaatataaa cattaaatac attacacgta gccacccgct
     1861 gtatagcata aaactaactt aaaattattc atatgcgttt cccggggccg aagcgttatt
     1921 gccgatattc ggtagccgat tccgctggct tttttttttt gaacgaggca aaaaagtttc
     1981 gtgactgaac ttcttcgtca ctggttttgt tgataatttt tatcgttttt tacttcgaga
     2041 ttgaaaacag tgttctgaat tttagcaacg atatcgcacc gcacgtttag cctgatcaaa
     2101 ccttatatat acctctacct taaaatatat atatatatag tattcgaatt ccgaattgtg
     2161 tgtctccgcg gttagtgatt gttatacaca aacagaatac aggaaagtaa atcaatacaa
     2221 taataaaagc tagcaaaatg acctatttcc cagaagaagt cgttgaacac atcttctctt
     2281 tcttgccagc tcaaagagat agaaatactg tttccttggt ttgtaaggtt tggtatgaaa
     2341 tcgaaagatt gagtagacgt ggtgttttcg tcggtaattg ttacgctgtt agagctggta
     2401 gagttgctgc tagattccca aatgttagag ccttgactgt caaaggtaaa ccacattttg
     2461 ctgactttaa cttggtccca ccagattggg gtggttatgc tggtccatgg attgaagctg
     2521 ccgctagagg ttgtcacggt ttggaagaat tgcgtatgaa aagaatggtc gtttctgacg
     2581 aatctttgga attattggcc cgttcttttc caagattcag agctttggtt ttgatctctt
     2641 gtgaaggttt cagtactgac ggtttggctg ccgttgcttc ccattgtaaa ttattgagag
     2701 aattggactt gcaagaaaat gaagtcgaag atagaggtcc aagatggtta tcttgttttc
     2761 cagactcttg tacttctttg gtctctttga acttcgcttg tattaagggt gaagtcaacg
     2821 ctggttcttt agaaagattg gtctcccgtt ctccaaactt acgttctttg agattgaatc
     2881 gttctgtttc cgttgatacc ttggctaaaa tcttattgag aaccccaaac ttggaagact
     2941 taggtactgg taacttgacc gacgatttcc aaactgaatc ctactttaaa ttaacttctg
     3001 ctttagaaaa gtgtaagatg ttgagatcct tatctggttt ttgggacgcc tctccagtct
     3061 gtttatcttt tatctatcca ttgtgtgctc aattaactgg tttaaactta tcttacgctc
     3121 caaccttgga tgctagtgac ttaaccaaga tgattagtag atgtgttaag ttacaaagat
     3181 tatgggtttt agattgtatt tccgataagg gtttacaagt cgttgctagt agctgtaagg
     3241 atttgcaaga attaagagtc tttccaagtg atttttacgt tgctggttac agcgctgtta
     3301 ctgaagaagg tttggtcgct gtttctttgg gttgtccaaa gttgaacagc ttattgtact
     3361 tttgtcatca aatgactaat gctgccttag tcactgttgc taagaactgt ccaaacttca
     3421 cccgtttcag attgtgtatt ttggaaccag gtaaaccaga cgttgtcacc tcccaaccat
     3481 tggatgaagg tttcggtgct atcgttagag aatgtaaggg tttgcaaaga ttatctatct
     3541 ctggtttgtt aactgacaaa gttttcatgt acatcggtaa atatgccaaa caattagaaa
     3601 tgttgagcat tgctttcgct ggtgactctg acaagggtat gatgcacgtt atgaatggtt
     3661 gtaagaattt gagaaagttg gaaattagag attctccatt cggtgacgct gccttattgg
     3721 gtaattttgc tagatacgaa actatgagaa gtttgtggat gagttcctgt aacgttacct
     3781 tgaaaggttg tcaagtctta gccagcaaga tgccaatgtt gaacgtcgaa gtcatcaacg
     3841 aaagagacgg tagtaacgaa atggaagaaa atcacggtga cttaccaaag gtagaaaagt
     3901 tgtacgtata cagaactact gctggtgcta gagatgatgc tccaaatttc gtaaagattt
     3961 tgtaactcga gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     4021 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     4081 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     4141 ttattacccg ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg
     4201 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6361 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6421 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6481 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6541 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6601 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6661 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6721 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6781 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat
     6841 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     6901 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     6961 agaacaaaaa cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga
     7021 gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag
     7081 accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa
     7141 acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt
     7201 tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt
     7261 gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt
     7321 taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt
     7381 gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg
     7441 cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt
     7501 gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga
     7561 ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa
     7621 ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat
     7681 atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa
     7741 agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta
     7801 tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg
     7861 gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg tattgggatc
     7921 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgctcga
     7981 ccagaaagcg accacccaac tttggctgat aatagcgtat aaacaatgca tactttgtac
     8041 gttcaaaata caatgcagta gatatattta tgcatattac atataataca tatcacatag
     8101 gaagcaacag gcgcgttgga cttttaattt tcgaggaccg cgaatcctta catcacaccc
     8161 aatcccccac aagtgatccc
//
