LOCUS       sequence-327148-        8183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of CUP1-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 8183)
  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 8183)
  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: pJAID5VY
FEATURES             Location/Qualifiers
     source          1..8183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             614..745
                     /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             755..1465
                     /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      1481..1751
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             2247..3974
                     /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(4204..4220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4204..4220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4217..4239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4228..4244
                     /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(4252..4258,4259..4276,4277..4282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4297..4318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4435..4452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4606..5194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4686..4705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5365..6156,6157..6225))
                     /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     5988..6007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6226..6330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6398..6416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6454..6476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6576..6595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6789..6811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6803..6820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6804..6820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7934..7967)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8178..8183,1..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 atgttcagcg aattaattaa cttccaaaat gaaggtcatg agtgccaatg
      481 ccaatgtggt agctgcaaaa ataatgaaca atgccaaaaa tcatgtagct gcccaacggg
      541 gtgtaacagc gacgacaaat gcccctgcgg taacaagtct gaagaaacca agaagtcatg
      601 ctgctctggg aaaccaaaag acccagctaa acctccagcc aaggcccaag tcgtcggctg
      661 gccaccagtt aggtcttaca gaaagaacgt aatggttagt tgtcaaaagt cctccggtgg
      721 tccagaagct gctgcctttg ttaagatggg atcctctaaa ggtgaagaat tattcactgg
      781 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
      841 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac
      901 tggtaaattg ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg
      961 ttttgcgaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     1021 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     1081 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     1141 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     1201 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa
     1261 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga
     1321 tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga
     1381 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     1441 ccatggtatg gatgaattgt acaaaagatc ttaaactagt attgaattga attgaaatcg
     1501 atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt
     1561 tattttttga atatttttta tttatatacg tatatataga ctattattta tcttttaatg
     1621 attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt
     1681 tttttcccat tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa
     1741 aattctgcgt tcgttaaagc tttcgagaag gatattattt gcggccgcga ttgcaagaat
     1801 tgatctccct catcgatgtc ataaatcagt aaatataaac attaaataca ttacacgtag
     1861 ccacccgctg tatagcataa aactaactta aaattattca tatgcgtttc ccggggccga
     1921 agcgttattg ccgatattcg gtagccgatt ccgctggctt tttttttttg aacgaggcaa
     1981 aaaagtttcg tgactgaact tcttcgtcac tggttttgtt gataattttt atcgtttttt
     2041 acttcgagat tgaaaacagt gttctgaatt ttagcaacga tatcgcaccg cacgtttagc
     2101 ctgatcaaac cttatatata cctctacctt aaaatatata tatatatagt attcgaattc
     2161 cgaattgtgt gtctccgcgg ttagtgattg ttatacacaa acagaataca ggaaagtaaa
     2221 tcaatacaat aataaattct agcaaaatga cctatttccc agaagaagtc gttgaacaca
     2281 tcttctcttt cttgccagct caaagagata gaaatactgt ttccttggtt tgtaaggttt
     2341 ggtatgaaat cgaaagattg agtagacgtg gtgttttcgt cggtaattgt tacgctgtta
     2401 gagctggtag agttgctgct agattcccaa atgttagagc cttgactgtc aaaggtaaac
     2461 cacattttgc tgactttaac ttggtcccac cagattgggg tggttatgct ggtccatgga
     2521 ttgaagctgc cgctagaggt tgtcacggtt tggaagaatt gcgtatgaaa agaatggtcg
     2581 tttctgacga atctttggaa ttattggccc gttcttttcc aagattcaga gctttggttt
     2641 tgatctcttg tgaaggtttc agtactgacg gtttggctgc cgttgcttcc cattgtaaat
     2701 tattgagaga attggacttg caagaaaatg aagtcgaaga tagaggtcca agatggttat
     2761 cttgttttcc agactcttgt acttctttgg tctctttgaa cttcgcttgt attaagggtg
     2821 aagtcaacgc tggttcttta gaaagattgg tctcccgttc tccaaactta cgttctttga
     2881 gattgaatcg ttctgtttcc gttgatacct tggctaaaat cttattgaga accccaaact
     2941 tggaagactt aggtactggt aacttgaccg acgatttcca aactgaatcc tactttaaat
     3001 taacttctgc tttagaaaag tgtaagatgt tgagatcctt atctggtttt tgggacgcct
     3061 ctccagtctg tttatctttt atctatccat tgtgtgctca attaactggt ttaaacttat
     3121 cttacgctcc aaccttggat gctagtgact taaccaagat gattagtaga tgtgttaagt
     3181 tacaaagatt atgggtttta gattgtattt ccgataaggg tttacaagtc gttgctagta
     3241 gctgtaagga tttgcaagaa ttaagagtct ttccaagtga tttttacgtt gctggttaca
     3301 gcgctgttac tgaagaaggt ttggtcgctg tttctttggg ttgtccaaag ttgaacagct
     3361 tattgtactt ttgtcatcaa atgactaatg ctgccttagt cactgttgct aagaactgtc
     3421 caaacttcac ccgtttcaga ttgtgtattt tggaaccagg taaaccagac gttgtcacct
     3481 cccaaccatt ggatgaaggt ttcggtgcta tcgttagaga atgtaagggt ttgcaaagat
     3541 tatctatctc tggtttgtta actgacaaag ttttcatgta catcggtaaa tatgccaaac
     3601 aattagaaat gttgagcatt gctttcgctg gtgactctga caagggtatg atgcacgtta
     3661 tgaatggttg taagaatttg agaaagttgg aaattagaga ttctccattc ggtgacgctg
     3721 ccttattggg taattttgct agatacgaaa ctatgagaag tttgtggatg agttcctgta
     3781 acgttacctt gaaaggttgt caagtcttag ccagcaagat gccaatgttg aacgtcgaag
     3841 tcatcaacga aagagacggt agtaacgaaa tggaagaaaa tcacggtgac ttaccaaagg
     3901 tagaaaagtt gtacgtatac agaactactg ctggtgctag agatgatgct ccaaatttcg
     3961 taaagatttt gtaaaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     4021 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     4081 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     4141 cagttattac ccgggaatct cggtcgtaat gatttaaatg catgcaagct tggcgtaatc
     4201 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4261 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4321 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4381 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4441 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4501 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5041 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5281 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5401 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5461 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5521 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5581 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5641 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5701 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5761 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5821 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5881 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5941 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6001 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6061 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6361 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6421 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6481 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6541 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6601 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6661 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6721 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6781 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcattt aaattttttt
     6841 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     6901 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     6961 cacagaacaa aaacctgcag gaaacgaaga taaaacatgt ccacaaaatc atataccagt
     7021 agagctgaga ctcatgcaag tccggttgca tcgaaacttt tacgtttaat ggatgaaaag
     7081 aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt gaaacttgtt
     7141 gaaacgttgg gtccatacat ttgccttttg aaaacacacg ttgatatctt ggatgatttc
     7201 agttatgagg gtactgtcgt tccattgaaa gcattggcag agaaatacaa gttcttgata
     7261 tttgaggaca gaaaattcgc cgatatcggt aacacagtca aattacaata tacatcgggc
     7321 gtttaccgta tcgcagaatg gtctgatatc accaacgccc acggggttac tggtgctggt
     7381 attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag gggattattg
     7441 atgcttgctg aattgtcttc caagggttct ctagcacacg gtgaatatac taagggtacc
     7501 gttgatattg caaagagtga taaagatttc gttattgggt tcattgctca gaacgatatg
     7561 ggaggaagag aagaagggtt tgattggcta atcatgaccc caggtgtagg tttagacgac
     7621 aaaggcgatg cattgggtca gcagtacaga accgtcgacg aagttgtaag tggtggatca
     7681 gatatcatca ttgttggcag aggacttttc gccaagggta gagatcctaa ggttgaaggt
     7741 gaaagataca gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc tccccattaa
     7801 ttatacagga aacttaatag aacaaatcac atatttaatc taatagccac ctgcattggc
     7861 acggtgcaac actacttcaa cttcatctta caaaaagatc acgtgatctg ttgtattggg
     7921 atctctagac ctaataactt cgtatagcat acattatacg aagttatatt aagggttgct
     7981 cgaccagaaa gcgaccaccc aactttggct gataatagcg tataaacaat gcatactttg
     8041 tacgttcaaa atacaatgca gtagatatat ttatgcatat tacatataat acatatcaca
     8101 taggaagcaa caggcgcgtt ggacttttaa ttttcgagga ccgcgaatcc ttacatcaca
     8161 cccaatcccc cacaagtgat ccc
//
