LOCUS       sequence-327679-        8360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of TRX1-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 8360)
  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 8360)
  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: pJAID6JY
FEATURES             Location/Qualifiers
     source          1..8360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             791..922
                     /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             932..1642
                     /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      1658..1928
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             2424..4151
                     /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(4381..4397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4381..4397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4394..4416)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4405..4421
                     /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(4429..4435,4436..4453,4454..4459))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4474..4495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4612..4629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4783..5371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4863..4882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5542..6333,6334..6402))
                     /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     6165..6184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6403..6507)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6575..6593
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6631..6653)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6753..6772
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6966..6988
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6980..6997
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6981..6997
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8111..8144)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8355..8360,1..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 atggttcaag cagtcgcatc taatctaagt tttaattaca aactcgagaa
      481 aatggttact caattcaaaa ctgccagcga attcgactct gcaattgctc aagacaagct
      541 agttgtcgta gatttctacg ccacttggtg cggtccatgt aaaatgattg ctccaatgat
      601 tgaaaaattc tctgaacaat acccacaagc tgatttctat aaattggatg tcgatgaatt
      661 gggtgatgtt gcacaaaaga atgaagtttc cgctatgcca actttgcttc tattcaagaa
      721 cggtaaggaa gttgcaaagg ttgttggtgc caacccagcg gctattaagc aagccattgc
      781 tgctaatgct ccaaaagacc cagctaaacc tccagccaag gcccaagtcg tcggctggcc
      841 accagttagg tcttacagaa agaacgtaat ggttagttgt caaaagtcct ccggtggtcc
      901 agaagctgct gcctttgtta agatgggatc ctctaaaggt gaagaattat tcactggtgt
      961 tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg
     1021 tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg
     1081 taaattgcca gttccatggc caaccttagt cactactttc ggttatggtg ttcaatgttt
     1141 tgcgagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg
     1201 ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga
     1261 agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa
     1321 agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta
     1381 catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat
     1441 tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg
     1501 tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat ccaaagatcc
     1561 aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca
     1621 tggtatggat gaattgtaca aaagatctta aactagtatt gaattgaatt gaaatcgata
     1681 gatcaatttt tttcttttct ctttccccat cctttacgct aaaataatag tttattttat
     1741 tttttgaata ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt
     1801 attaagattt ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt
     1861 ttcccattcg atatttctat gttcgggttc agcgtatttt aagtttaata actcgaaaat
     1921 tctgcgttcg ttaaagcttt cgagaaggat attatttgcg gccgcgattg caagaattga
     1981 tctccctcat cgatgtcata aatcagtaaa tataaacatt aaatacatta cacgtagcca
     2041 cccgctgtat agcataaaac taacttaaaa ttattcatat gcgtttcccg gggccgaagc
     2101 gttattgccg atattcggta gccgattccg ctggcttttt ttttttgaac gaggcaaaaa
     2161 agtttcgtga ctgaacttct tcgtcactgg ttttgttgat aatttttatc gttttttact
     2221 tcgagattga aaacagtgtt ctgaatttta gcaacgatat cgcaccgcac gtttagcctg
     2281 atcaaacctt atatatacct ctaccttaaa atatatatat atatagtatt cgaattccga
     2341 attgtgtgtc tccgcggtta gtgattgtta tacacaaaca gaatacagga aagtaaatca
     2401 atacaataat aaattctagc aaaatgacct atttcccaga agaagtcgtt gaacacatct
     2461 tctctttctt gccagctcaa agagatagaa atactgtttc cttggtttgt aaggtttggt
     2521 atgaaatcga aagattgagt agacgtggtg ttttcgtcgg taattgttac gctgttagag
     2581 ctggtagagt tgctgctaga ttcccaaatg ttagagcctt gactgtcaaa ggtaaaccac
     2641 attttgctga ctttaacttg gtcccaccag attggggtgg ttatgctggt ccatggattg
     2701 aagctgccgc tagaggttgt cacggtttgg aagaattgcg tatgaaaaga atggtcgttt
     2761 ctgacgaatc tttggaatta ttggcccgtt cttttccaag attcagagct ttggttttga
     2821 tctcttgtga aggtttcagt actgacggtt tggctgccgt tgcttcccat tgtaaattat
     2881 tgagagaatt ggacttgcaa gaaaatgaag tcgaagatag aggtccaaga tggttatctt
     2941 gttttccaga ctcttgtact tctttggtct ctttgaactt cgcttgtatt aagggtgaag
     3001 tcaacgctgg ttctttagaa agattggtct cccgttctcc aaacttacgt tctttgagat
     3061 tgaatcgttc tgtttccgtt gataccttgg ctaaaatctt attgagaacc ccaaacttgg
     3121 aagacttagg tactggtaac ttgaccgacg atttccaaac tgaatcctac tttaaattaa
     3181 cttctgcttt agaaaagtgt aagatgttga gatccttatc tggtttttgg gacgcctctc
     3241 cagtctgttt atcttttatc tatccattgt gtgctcaatt aactggttta aacttatctt
     3301 acgctccaac cttggatgct agtgacttaa ccaagatgat tagtagatgt gttaagttac
     3361 aaagattatg ggttttagat tgtatttccg ataagggttt acaagtcgtt gctagtagct
     3421 gtaaggattt gcaagaatta agagtctttc caagtgattt ttacgttgct ggttacagcg
     3481 ctgttactga agaaggtttg gtcgctgttt ctttgggttg tccaaagttg aacagcttat
     3541 tgtacttttg tcatcaaatg actaatgctg ccttagtcac tgttgctaag aactgtccaa
     3601 acttcacccg tttcagattg tgtattttgg aaccaggtaa accagacgtt gtcacctccc
     3661 aaccattgga tgaaggtttc ggtgctatcg ttagagaatg taagggtttg caaagattat
     3721 ctatctctgg tttgttaact gacaaagttt tcatgtacat cggtaaatat gccaaacaat
     3781 tagaaatgtt gagcattgct ttcgctggtg actctgacaa gggtatgatg cacgttatga
     3841 atggttgtaa gaatttgaga aagttggaaa ttagagattc tccattcggt gacgctgcct
     3901 tattgggtaa ttttgctaga tacgaaacta tgagaagttt gtggatgagt tcctgtaacg
     3961 ttaccttgaa aggttgtcaa gtcttagcca gcaagatgcc aatgttgaac gtcgaagtca
     4021 tcaacgaaag agacggtagt aacgaaatgg aagaaaatca cggtgactta ccaaaggtag
     4081 aaaagttgta cgtatacaga actactgctg gtgctagaga tgatgctcca aatttcgtaa
     4141 agattttgta aaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     4201 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     4261 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     4321 ttattacccg ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg
     4381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6541 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6601 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6661 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6721 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6781 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6841 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6901 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6961 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat
     7021 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     7081 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     7141 agaacaaaaa cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga
     7201 gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag
     7261 accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa
     7321 acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt
     7381 tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt
     7441 gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt
     7501 taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt
     7561 gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg
     7621 cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt
     7681 gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga
     7741 ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa
     7801 ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat
     7861 atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa
     7921 agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta
     7981 tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg
     8041 gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg tattgggatc
     8101 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgctcga
     8161 ccagaaagcg accacccaac tttggctgat aatagcgtat aaacaatgca tactttgtac
     8221 gttcaaaata caatgcagta gatatattta tgcatattac atataataca tatcacatag
     8281 gaagcaacag gcgcgttgga cttttaattt tcgaggaccg cgaatcctta catcacaccc
     8341 aatcccccac aagtgatccc
//
