LOCUS       sequence-327150-        8495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of yEGFP and SKP1-fusing rice auxin receptor 
            gene OsTIR1. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  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 8495)
  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: pJAID8Z
FEATURES             Location/Qualifiers
     source          1..8495
                     /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"
     protein_bind    2127..2138
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             complement(2524..2541)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             2553..4280
                     /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(4516..4532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4529..4551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4540..4556
                     /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(4564..4570,4571..4588,4589..4594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4609..4630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4747..4764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4918..5506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4998..5017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5677..6468,6469..6537))
                     /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     6300..6319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6538..6642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6710..6728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6766..6788)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6888..6907
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7101..7123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7115..7132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7116..7132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8246..8279)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8490..8495,1..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 aaatggtgac ttctaatgtt gtcctagtga gtggtgaggg tgaacgattc accgtagaca
     1981 agaaaatcgc ggagagatcc ctgctattga aaaactatct gaacgatatg catgacagca
     2041 acctacaaaa caactcagat tcagaatcag attcagactc ggaaacgaac cacaaaagca
     2101 aagacaacaa taacggtgac gacgacgatg aggatgacga cgagattgtg atgcccgtgc
     2161 ccaatgttcg gtcttccgtt ttgcagaagg ttatcgaatg ggccgagcac cacagagact
     2221 ctaacttccc tgacgaagat gacgacgatt ccagaaagag tgctcccgtg gactcatggg
     2281 acagagaatt cctgaaagtg gaccaagaga tgctatacga aattattctg gcggcaaact
     2341 acctcaacat caagcccttg cttgacgctg gctgcaaggt ggttgctgaa atgatcaggg
     2401 gaaggtctcc cgaggaaatt agaagaacgt ttaatatcgt caacgacttc actccagagg
     2461 aggaggctgc cattaggcgt gaaaatgaat gggctgaaga ccgtggtatc tttgatttgg
     2521 gtgctggtgc tggtgctggt gaccttactt ctatgaccta tttcccagaa gaagtcgttg
     2581 aacacatctt ctctttcttg ccagctcaaa gagatagaaa tactgtttcc ttggtttgta
     2641 aggtttggta tgaaatcgaa agattgagta gacgtggtgt tttcgtcggt aattgttacg
     2701 ctgttagagc tggtagagtt gctgctagat tcccaaatgt tagagccttg actgtcaaag
     2761 gtaaaccaca ttttgctgac tttaacttgg tcccaccaga ttggggtggt tatgctggtc
     2821 catggattga agctgccgct agaggttgtc acggtttgga agaattgcgt atgaaaagaa
     2881 tggtcgtttc tgacgaatct ttggaattat tggcccgttc ttttccaaga ttcagagctt
     2941 tggttttgat ctcttgtgaa ggtttcagta ctgacggttt ggctgccgtt gcttcccatt
     3001 gtaaattatt gagagaattg gacttgcaag aaaatgaagt cgaagataga ggtccaagat
     3061 ggttatcttg ttttccagac tcttgtactt ctttggtctc tttgaacttc gcttgtatta
     3121 agggtgaagt caacgctggt tctttagaaa gattggtctc ccgttctcca aacttacgtt
     3181 ctttgagatt gaatcgttct gtttccgttg ataccttggc taaaatctta ttgagaaccc
     3241 caaacttgga agacttaggt actggtaact tgaccgacga tttccaaact gaatcctact
     3301 ttaaattaac ttctgcttta gaaaagtgta agatgttgag atccttatct ggtttttggg
     3361 acgcctctcc agtctgttta tcttttatct atccattgtg tgctcaatta actggtttaa
     3421 acttatctta cgctccaacc ttggatgcta gtgacttaac caagatgatt agtagatgtg
     3481 ttaagttaca aagattatgg gttttagatt gtatttccga taagggttta caagtcgttg
     3541 ctagtagctg taaggatttg caagaattaa gagtctttcc aagtgatttt tacgttgctg
     3601 gttacagcgc tgttactgaa gaaggtttgg tcgctgtttc tttgggttgt ccaaagttga
     3661 acagcttatt gtacttttgt catcaaatga ctaatgctgc cttagtcact gttgctaaga
     3721 actgtccaaa cttcacccgt ttcagattgt gtattttgga accaggtaaa ccagacgttg
     3781 tcacctccca accattggat gaaggtttcg gtgctatcgt tagagaatgt aagggtttgc
     3841 aaagattatc tatctctggt ttgttaactg acaaagtttt catgtacatc ggtaaatatg
     3901 ccaaacaatt agaaatgttg agcattgctt tcgctggtga ctctgacaag ggtatgatgc
     3961 acgttatgaa tggttgtaag aatttgagaa agttggaaat tagagattct ccattcggtg
     4021 acgctgcctt attgggtaat tttgctagat acgaaactat gagaagtttg tggatgagtt
     4081 cctgtaacgt taccttgaaa ggttgtcaag tcttagccag caagatgcca atgttgaacg
     4141 tcgaagtcat caacgaaaga gacggtagta acgaaatgga agaaaatcac ggtgacttac
     4201 caaaggtaga aaagttgtac gtatacagaa ctactgctgg tgctagagat gatgctccaa
     4261 atttcgtaaa gattttgtaa ctcgagagga ctatttgcaa agggaaggga tgctaaggta
     4321 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     4381 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     4441 atttaattat atcagttatt acccgggaat ctcggtcgta atgatttaaa tgcatgcaag
     4501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6661 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6721 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6781 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6841 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6901 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6961 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7021 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7081 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcat
     7141 ttaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     7201 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     7261 ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaaacat gtccacaaaa
     7321 tcatatacca gtagagctga gactcatgca agtccggttg catcgaaact tttacgttta
     7381 atggatgaaa agaagaccaa tttgtgtgct tctcttgacg ttcgttcgac tgatgagcta
     7441 ttgaaacttg ttgaaacgtt gggtccatac atttgccttt tgaaaacaca cgttgatatc
     7501 ttggatgatt tcagttatga gggtactgtc gttccattga aagcattggc agagaaatac
     7561 aagttcttga tatttgagga cagaaaattc gccgatatcg gtaacacagt caaattacaa
     7621 tatacatcgg gcgtttaccg tatcgcagaa tggtctgata tcaccaacgc ccacggggtt
     7681 actggtgctg gtattgttgc tggcttgaaa caaggtgcgc aagaggtcac caaagaacca
     7741 aggggattat tgatgcttgc tgaattgtct tccaagggtt ctctagcaca cggtgaatat
     7801 actaagggta ccgttgatat tgcaaagagt gataaagatt tcgttattgg gttcattgct
     7861 cagaacgata tgggaggaag agaagaaggg tttgattggc taatcatgac cccaggtgta
     7921 ggtttagacg acaaaggcga tgcattgggt cagcagtaca gaaccgtcga cgaagttgta
     7981 agtggtggat cagatatcat cattgttggc agaggacttt tcgccaaggg tagagatcct
     8041 aaggttgaag gtgaaagata cagaaatgct ggatgggaag cgtaccaaaa gagaatcagc
     8101 gctccccatt aattatacag gaaacttaat agaacaaatc acatatttaa tctaatagcc
     8161 acctgcattg gcacggtgca acactacttc aacttcatct tacaaaaaga tcacgtgatc
     8221 tgttgtattg ggatctctag acctaataac ttcgtatagc atacattata cgaagttata
     8281 ttaagggttg ctcgaccaga aagcgaccac ccaactttgg ctgataatag cgtataaaca
     8341 atgcatactt tgtacgttca aaatacaatg cagtagatat atttatgcat attacatata
     8401 atacatatca cataggaagc aacaggcgcg ttggactttt aattttcgag gaccgcgaat
     8461 ccttacatca cacccaatcc cccacaagtg atccc
//
