LOCUS       sequence-327154-        8210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of nucleus-targeting 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 8210)
  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 8210)
  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: pJAID9L105
FEATURES             Location/Qualifiers
     source          1..8210
                     /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"
     CDS             1472..1492
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1502..1772
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             2268..3995
                     /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(4231..4247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4244..4266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4255..4271
                     /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(4279..4285,4286..4303,4304..4309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4324..4345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4462..4479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4633..5221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4713..4732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5392..6183,6184..6252))
                     /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     6015..6034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6253..6357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6425..6443
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6481..6503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6603..6622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6816..6838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6830..6847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6831..6847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7961..7994)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8205..8210,1..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 aatgtacccc acctaagaag aaaagaaagg tttaaactag
     1501 tattgaattg aattgaaatc gatagatcaa tttttttctt ttctctttcc ccatccttta
     1561 cgctaaaata atagtttatt ttattttttg aatatttttt atttatatac gtatatatag
     1621 actattattt atcttttaat gattattaag atttttatta aaaaaaaatt cgctcctctt
     1681 ttaatgcctt tatgcagttt ttttttccca ttcgatattt ctatgttcgg gttcagcgta
     1741 ttttaagttt aataactcga aaattctgcg ttcgttaaag ctttcgagaa ggatattatt
     1801 tgcggccgcg attgcaagaa ttgatctccc tcatcgatgt cataaatcag taaatataaa
     1861 cattaaatac attacacgta gccacccgct gtatagcata aaactaactt aaaattattc
     1921 atatgcgttt cccggggccg aagcgttatt gccgatattc ggtagccgat tccgctggct
     1981 tttttttttt gaacgaggca aaaaagtttc gtgactgaac ttcttcgtca ctggttttgt
     2041 tgataatttt tatcgttttt tacttcgaga ttgaaaacag tgttctgaat tttagcaacg
     2101 atatcgcacc gcacgtttag cctgatcaaa ccttatatat acctctacct taaaatatat
     2161 atatatatag tattcgaatt ccgaattgtg tgtctccgcg gttagtgatt gttatacaca
     2221 aacagaatac aggaaagtaa atcaatacaa taataaaagc tagcaaaatg acctatttcc
     2281 cagaagaagt cgttgaacac atcttctctt tcttgccagc tcaaagagat agaaatactg
     2341 tttccttggt ttgtaaggtt tggtatgaaa tcgaaagatt gagtagacgt ggtgttttcg
     2401 tcggtaattg ttacgctgtt agagctggta gagttgctgc tagattccca aatgttagag
     2461 ccttgactgt caaaggtaaa ccacattttg ctgactttaa cttggtccca ccagattggg
     2521 gtggttatgc tggtccatgg attgaagctg ccgctagagg ttgtcacggt ttggaagaat
     2581 tgcgtatgaa aagaatggtc gtttctgacg aatctttgga attattggcc cgttcttttc
     2641 caagattcag agctttggtt ttgatctctt gtgaaggttt cagtactgac ggtttggctg
     2701 ccgttgcttc ccattgtaaa ttattgagag aattggactt gcaagaaaat gaagtcgaag
     2761 atagaggtcc aagatggtta tcttgttttc cagactcttg tacttctttg gtctctttga
     2821 acttcgcttg tattaagggt gaagtcaacg ctggttcttt agaaagattg gtctcccgtt
     2881 ctccaaactt acgttctttg agattgaatc gttctgtttc cgttgatacc ttggctaaaa
     2941 tcttattgag aaccccaaac ttggaagact taggtactgg taacttgacc gacgatttcc
     3001 aaactgaatc ctactttaaa ttaacttctg ctttagaaaa gtgtaagatg ttgagatcct
     3061 tatctggttt ttgggacgcc tctccagtct gtttatcttt tatctatcca ttgtgtgctc
     3121 aattaactgg tttaaactta tcttacgctc caaccttgga tgctagtgac ttaaccaaga
     3181 tgattagtag atgtgttaag ttacaaagat tatgggtttt agattgtatt tccgataagg
     3241 gtttacaagt cgttgctagt agctgtaagg atttgcaaga attaagagtc tttccaagtg
     3301 atttttacgt tgctggttac agcgctgtta ctgaagaagg tttggtcgct gtttctttgg
     3361 gttgtccaaa gttgaacagc ttattgtact tttgtcatca aatgactaat gctgccttag
     3421 tcactgttgc taagaactgt ccaaacttca cccgtttcag attgtgtatt ttggaaccag
     3481 gtaaaccaga cgttgtcacc tcccaaccat tggatgaagg tttcggtgct atcgttagag
     3541 aatgtaaggg tttgcaaaga ttatctatct ctggtttgtt aactgacaaa gttttcatgt
     3601 acatcggtaa atatgccaaa caattagaaa tgttgagcat tgctttcgct ggtgactctg
     3661 acaagggtat gatgcacgtt atgaatggtt gtaagaattt gagaaagttg gaaattagag
     3721 attctccatt cggtgacgct gccttattgg gtaattttgc tagatacgaa actatgagaa
     3781 gtttgtggat gagttcctgt aacgttacct tgaaaggttg tcaagtctta gccagcaaga
     3841 tgccaatgtt gaacgtcgaa gtcatcaacg aaagagacgg tagtaacgaa atggaagaaa
     3901 atcacggtga cttaccaaag gtagaaaagt tgtacgtata cagaactact gctggtgcta
     3961 gagatgatgc tccaaatttc gtaaagattt tgtaactcga gaggactatt tgcaaaggga
     4021 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     4081 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     4141 caaattagag cttcaattta attatatcag ttattacccg ggaatctcgg tcgtaatgat
     4201 ttaaatgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6361 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6421 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     6481 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6541 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     6601 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6661 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     6721 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6781 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6841 ggccagtgaa ttcatttaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     6901 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     6961 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     7021 aacatgtcca caaaatcata taccagtaga gctgagactc atgcaagtcc ggttgcatcg
     7081 aaacttttac gtttaatgga tgaaaagaag accaatttgt gtgcttctct tgacgttcgt
     7141 tcgactgatg agctattgaa acttgttgaa acgttgggtc catacatttg ccttttgaaa
     7201 acacacgttg atatcttgga tgatttcagt tatgagggta ctgtcgttcc attgaaagca
     7261 ttggcagaga aatacaagtt cttgatattt gaggacagaa aattcgccga tatcggtaac
     7321 acagtcaaat tacaatatac atcgggcgtt taccgtatcg cagaatggtc tgatatcacc
     7381 aacgcccacg gggttactgg tgctggtatt gttgctggct tgaaacaagg tgcgcaagag
     7441 gtcaccaaag aaccaagggg attattgatg cttgctgaat tgtcttccaa gggttctcta
     7501 gcacacggtg aatatactaa gggtaccgtt gatattgcaa agagtgataa agatttcgtt
     7561 attgggttca ttgctcagaa cgatatggga ggaagagaag aagggtttga ttggctaatc
     7621 atgaccccag gtgtaggttt agacgacaaa ggcgatgcat tgggtcagca gtacagaacc
     7681 gtcgacgaag ttgtaagtgg tggatcagat atcatcattg ttggcagagg acttttcgcc
     7741 aagggtagag atcctaaggt tgaaggtgaa agatacagaa atgctggatg ggaagcgtac
     7801 caaaagagaa tcagcgctcc ccattaatta tacaggaaac ttaatagaac aaatcacata
     7861 tttaatctaa tagccacctg cattggcacg gtgcaacact acttcaactt catcttacaa
     7921 aaagatcacg tgatctgttg tattgggatc tctagaccta ataacttcgt atagcataca
     7981 ttatacgaag ttatattaag ggttgctcga ccagaaagcg accacccaac tttggctgat
     8041 aatagcgtat aaacaatgca tactttgtac gttcaaaata caatgcagta gatatattta
     8101 tgcatattac atataataca tatcacatag gaagcaacag gcgcgttgga cttttaattt
     8161 tcgaggaccg cgaatcctta catcacaccc aatcccccac aagtgatccc
//
