LOCUS       sequence-327177-        7085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative expression of SKP1-fusing rice auxin receptor gene 
            OsTIR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7085)
  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 7085)
  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: pISKP1-TIR1
FEATURES             Location/Qualifiers
     source          1..7085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2..272
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     protein_bind    972..983
                     /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(1369..1386)
                     /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             1398..3125
                     /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(3361..3377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3361..3377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3374..3396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3385..3401
                     /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(3409..3415,3416..3433,3434..3439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3454..3475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3592..3609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3763..4351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3843..3862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4522..5313,5314..5382))
                     /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     5145..5164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5383..5487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5555..5573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5611..5633)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5733..5752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5946..5968
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5960..5977
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5961..5977
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tattgaattg aattgaaatc gatagatcaa tttttttctt ttctctttcc ccatccttta
       61 cgctaaaata atagtttatt ttattttttg aatatttttt atttatatac gtatatatag
      121 actattattt atcttttaat gattattaag atttttatta aaaaaaaatt cgctcctctt
      181 ttaatgcctt tatgcagttt ttttttccca ttcgatattt ctatgttcgg gttcagcgta
      241 ttttaagttt aataactcga aaattctgcg ttcgttaaag ctttcgagaa ggatattatt
      301 tgcggccgcg attgcaagaa ttgatctccc tcatcgatgt cataaatcag taaatataaa
      361 cattaaatac attacacgta gccacccgct gtatagcata aaactaactt aaaattattc
      421 atatgcgttt cccggggccg aagcgttatt gccgatattc ggtagccgat tccgctggct
      481 tttttttttt gaacgaggca aaaaagtttc gtgactgaac ttcttcgtca ctggttttgt
      541 tgataatttt tatcgttttt tacttcgaga ttgaaaacag tgttctgaat tttagcaacg
      601 atatcgcacc gcacgtttag cctgatcaaa ccttatatat acctctacct taaaatatat
      661 atatatatag tattcgaatt ccgaattgtg tgtctccgcg gttagtgatt gttatacaca
      721 aacagaatac aggaaagtaa atcaatacaa taataaaagc tagcaaaatg gtgacttcta
      781 atgttgtcct agtgagtggt gagggtgaac gattcaccgt agacaagaaa atcgcggaga
      841 gatccctgct attgaaaaac tatctgaacg atatgcatga cagcaaccta caaaacaact
      901 cagattcaga atcagattca gactcggaaa cgaaccacaa aagcaaagac aacaataacg
      961 gtgacgacga cgatgaggat gacgacgaga ttgtgatgcc cgtgcccaat gttcggtctt
     1021 ccgttttgca gaaggttatc gaatgggccg agcaccacag agactctaac ttccctgacg
     1081 aagatgacga cgattccaga aagagtgctc ccgtggactc atgggacaga gaattcctga
     1141 aagtggacca agagatgcta tacgaaatta ttctggcggc aaactacctc aacatcaagc
     1201 ccttgcttga cgctggctgc aaggtggttg ctgaaatgat caggggaagg tctcccgagg
     1261 aaattagaag aacgtttaat atcgtcaacg acttcactcc agaggaggag gctgccatta
     1321 ggcgtgaaaa tgaatgggct gaagaccgtg gtatctttga tttgggtgct ggtgctggtg
     1381 ctggtgacct tacttctatg acctatttcc cagaagaagt cgttgaacac atcttctctt
     1441 tcttgccagc tcaaagagat agaaatactg tttccttggt ttgtaaggtt tggtatgaaa
     1501 tcgaaagatt gagtagacgt ggtgttttcg tcggtaattg ttacgctgtt agagctggta
     1561 gagttgctgc tagattccca aatgttagag ccttgactgt caaaggtaaa ccacattttg
     1621 ctgactttaa cttggtccca ccagattggg gtggttatgc tggtccatgg attgaagctg
     1681 ccgctagagg ttgtcacggt ttggaagaat tgcgtatgaa aagaatggtc gtttctgacg
     1741 aatctttgga attattggcc cgttcttttc caagattcag agctttggtt ttgatctctt
     1801 gtgaaggttt cagtactgac ggtttggctg ccgttgcttc ccattgtaaa ttattgagag
     1861 aattggactt gcaagaaaat gaagtcgaag atagaggtcc aagatggtta tcttgttttc
     1921 cagactcttg tacttctttg gtctctttga acttcgcttg tattaagggt gaagtcaacg
     1981 ctggttcttt agaaagattg gtctcccgtt ctccaaactt acgttctttg agattgaatc
     2041 gttctgtttc cgttgatacc ttggctaaaa tcttattgag aaccccaaac ttggaagact
     2101 taggtactgg taacttgacc gacgatttcc aaactgaatc ctactttaaa ttaacttctg
     2161 ctttagaaaa gtgtaagatg ttgagatcct tatctggttt ttgggacgcc tctccagtct
     2221 gtttatcttt tatctatcca ttgtgtgctc aattaactgg tttaaactta tcttacgctc
     2281 caaccttgga tgctagtgac ttaaccaaga tgattagtag atgtgttaag ttacaaagat
     2341 tatgggtttt agattgtatt tccgataagg gtttacaagt cgttgctagt agctgtaagg
     2401 atttgcaaga attaagagtc tttccaagtg atttttacgt tgctggttac agcgctgtta
     2461 ctgaagaagg tttggtcgct gtttctttgg gttgtccaaa gttgaacagc ttattgtact
     2521 tttgtcatca aatgactaat gctgccttag tcactgttgc taagaactgt ccaaacttca
     2581 cccgtttcag attgtgtatt ttggaaccag gtaaaccaga cgttgtcacc tcccaaccat
     2641 tggatgaagg tttcggtgct atcgttagag aatgtaaggg tttgcaaaga ttatctatct
     2701 ctggtttgtt aactgacaaa gttttcatgt acatcggtaa atatgccaaa caattagaaa
     2761 tgttgagcat tgctttcgct ggtgactctg acaagggtat gatgcacgtt atgaatggtt
     2821 gtaagaattt gagaaagttg gaaattagag attctccatt cggtgacgct gccttattgg
     2881 gtaattttgc tagatacgaa actatgagaa gtttgtggat gagttcctgt aacgttacct
     2941 tgaaaggttg tcaagtctta gccagcaaga tgccaatgtt gaacgtcgaa gtcatcaacg
     3001 aaagagacgg tagtaacgaa atggaagaaa atcacggtga cttaccaaag gtagaaaagt
     3061 tgtacgtata cagaactact gctggtgcta gagatgatgc tccaaatttc gtaaagattt
     3121 tgtaactcga gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     3181 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     3241 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     3301 ttattacccg ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg
     3361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5521 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5581 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5641 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5701 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5761 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5821 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5881 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5941 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat
     6001 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     6061 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     6121 agaacaaaaa cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga
     6181 gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag
     6241 accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa
     6301 acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt
     6361 tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt
     6421 gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt
     6481 taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt
     6541 gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg
     6601 cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt
     6661 gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga
     6721 ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa
     6781 ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat
     6841 atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa
     6901 agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta
     6961 tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg
     7021 gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg tattgggatc
     7081 tctag
//
