LOCUS       sequence-327146-        7868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of yEGFP and rice auxin receptor gene OsTIR1.
            OsTIR1 is codon-optimised for expression in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7868)
  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 7868)
  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: pJAID6A
FEATURES             Location/Qualifiers
     source          1..7868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..1736
                     /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(1966..1982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1966..1982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1979..2001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1990..2006
                     /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(2014..2020,2021..2038,2039..2044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2059..2080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2197..2214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2368..2956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2448..2467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3127..3918,3919..3987))
                     /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     3750..3769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3988..4092)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4160..4178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4216..4238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4338..4357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4551..4573
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4565..4582
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4566..4582
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5696..5729)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5940..6353
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6385..7095
                     /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      7111..7381
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
ORIGIN
        1 ctagcaaaat gacctatttc ccagaagaag tcgttgaaca catcttctct ttcttgccag
       61 ctcaaagaga tagaaatact gtttccttgg tttgtaaggt ttggtatgaa atcgaaagat
      121 tgagtagacg tggtgttttc gtcggtaatt gttacgctgt tagagctggt agagttgctg
      181 ctagattccc aaatgttaga gccttgactg tcaaaggtaa accacatttt gctgacttta
      241 acttggtccc accagattgg ggtggttatg ctggtccatg gattgaagct gccgctagag
      301 gttgtcacgg tttggaagaa ttgcgtatga aaagaatggt cgtttctgac gaatctttgg
      361 aattattggc ccgttctttt ccaagattca gagctttggt tttgatctct tgtgaaggtt
      421 tcagtactga cggtttggct gccgttgctt cccattgtaa attattgaga gaattggact
      481 tgcaagaaaa tgaagtcgaa gatagaggtc caagatggtt atcttgtttt ccagactctt
      541 gtacttcttt ggtctctttg aacttcgctt gtattaaggg tgaagtcaac gctggttctt
      601 tagaaagatt ggtctcccgt tctccaaact tacgttcttt gagattgaat cgttctgttt
      661 ccgttgatac cttggctaaa atcttattga gaaccccaaa cttggaagac ttaggtactg
      721 gtaacttgac cgacgatttc caaactgaat cctactttaa attaacttct gctttagaaa
      781 agtgtaagat gttgagatcc ttatctggtt tttgggacgc ctctccagtc tgtttatctt
      841 ttatctatcc attgtgtgct caattaactg gtttaaactt atcttacgct ccaaccttgg
      901 atgctagtga cttaaccaag atgattagta gatgtgttaa gttacaaaga ttatgggttt
      961 tagattgtat ttccgataag ggtttacaag tcgttgctag tagctgtaag gatttgcaag
     1021 aattaagagt ctttccaagt gatttttacg ttgctggtta cagcgctgtt actgaagaag
     1081 gtttggtcgc tgtttctttg ggttgtccaa agttgaacag cttattgtac ttttgtcatc
     1141 aaatgactaa tgctgcctta gtcactgttg ctaagaactg tccaaacttc acccgtttca
     1201 gattgtgtat tttggaacca ggtaaaccag acgttgtcac ctcccaacca ttggatgaag
     1261 gtttcggtgc tatcgttaga gaatgtaagg gtttgcaaag attatctatc tctggtttgt
     1321 taactgacaa agttttcatg tacatcggta aatatgccaa acaattagaa atgttgagca
     1381 ttgctttcgc tggtgactct gacaagggta tgatgcacgt tatgaatggt tgtaagaatt
     1441 tgagaaagtt ggaaattaga gattctccat tcggtgacgc tgccttattg ggtaattttg
     1501 ctagatacga aactatgaga agtttgtgga tgagttcctg taacgttacc ttgaaaggtt
     1561 gtcaagtctt agccagcaag atgccaatgt tgaacgtcga agtcatcaac gaaagagacg
     1621 gtagtaacga aatggaagaa aatcacggtg acttaccaaa ggtagaaaag ttgtacgtat
     1681 acagaactac tgctggtgct agagatgatg ctccaaattt cgtaaagatt ttgtaaagga
     1741 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     1801 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     1861 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat
     1921 ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     1981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4141 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4201 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4261 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4321 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4381 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     4441 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4501 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4561 acgttgtaaa acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa
     4621 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     4681 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc
     4741 aggaaacgaa gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca
     4801 agtccggttg catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct
     4861 tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac
     4921 atttgccttt tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc
     4981 gttccattga aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc
     5041 gccgatatcg gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa
     5101 tggtctgata tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa
     5161 caaggtgcgc aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct
     5221 tccaagggtt ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt
     5281 gataaagatt tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg
     5341 tttgattggc taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt
     5401 cagcagtaca gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc
     5461 agaggacttt tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct
     5521 ggatgggaag cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat
     5581 agaacaaatc acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc
     5641 aacttcatct tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac
     5701 ttcgtatagc atacattata cgaagttata ttaagggttg ctcgaccaga aagcgaccac
     5761 ccaactttgg ctgataatag cgtataaaca atgcatactt tgtacgttca aaatacaatg
     5821 cagtagatat atttatgcat attacatata atacatatca cataggaagc aacaggcgcg
     5881 ttggactttt aattttcgag gaccgcgaat ccttacatca cacccaatcc cccacaagtg
     5941 atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc
     6001 tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc
     6061 cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa
     6121 gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc
     6181 tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt
     6241 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac
     6301 aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat
     6361 tacaaactcg agaaaatggg atcctctaaa ggtgaagaat tattcactgg tgttgtccca
     6421 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt
     6481 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg
     6541 ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgcgaga
     6601 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt
     6661 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag
     6721 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat
     6781 ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg
     6841 gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat
     6901 ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc
     6961 ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa
     7021 aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg
     7081 gatgaattgt acaaaagatc ttaaactagt attgaattga attgaaatcg atagatcaat
     7141 ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt tattttttga
     7201 atatttttta tttatatacg tatatataga ctattattta tcttttaatg attattaaga
     7261 tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt tttttcccat
     7321 tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa aattctgcgt
     7381 tcgttaaagc tttcgagaag gatattattt gcggccgcga ttgcaagaat tgatctccct
     7441 catcgatgtc ataaatcagt aaatataaac attaaataca ttacacgtag ccacccgctg
     7501 tatagcataa aactaactta aaattattca tatgcgtttc ccggggccga agcgttattg
     7561 ccgatattcg gtagccgatt ccgctggctt tttttttttg aacgaggcaa aaaagtttcg
     7621 tgactgaact tcttcgtcac tggttttgtt gataattttt atcgtttttt acttcgagat
     7681 tgaaaacagt gttctgaatt ttagcaacga tatcgcaccg cacgtttagc ctgatcaaac
     7741 cttatatata cctctacctt aaaatatata tatatatagt attcgaattc cgaattgtgt
     7801 gtctccgcgg ttagtgattg ttatacacaa acagaataca ggaaagtaaa tcaatacaat
     7861 aataaatt
//
