LOCUS       sequence-370256-        7622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabidopsis Nuclear Auxin Signaling Receptor TIR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  AUTHORS   Ramos Baez R, Buckley Y, Yu H, Chen Z, Gallavotti A, Nemhauser JL, 
            Moss BL
  TITLE     A Synthetic Approach Allows Rapid Characterization of the Maize 
            Nuclear Auxin Response Circuit.
  JOURNAL   Plant Physiol. 2020 Apr;182(4):1713-1722. doi: 10.1104/pp.19.01475. 
            Epub 2020 Mar 2.
  PUBMED    32123041
REFERENCE   2  (bases 1 to 7622)
  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"; doi: 
            "10.1104/pp.19.01475"; journalName: "Plant Physiol"; date: 
            "2020-04"; volume: "182"; issue: "4"; pages: "1713-1722"
COMMENT     Sequence Label: pGP8GF-AtTIR1
FEATURES             Location/Qualifiers
     source          1..7622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      659..824
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(714..738)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        847..1490
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1384..1407
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1496..1512
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1496..1512
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3319..3384
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3385..3402
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3450..3574)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3607..3623)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3608..3624)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3626..3872
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3642..3660)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4886..4902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4886..4902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4899..4921)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4910..4926
                     /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(4934..4940,4941..4958,4959..4964))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4979..5000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5117..5134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5288..5876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5368..5387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6047..6838,6839..6907))
                     /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     6670..6689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6908..7012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7080..7098
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7136..7158)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7258..7277
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7430..7617
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(7618..7622,1..658)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
ORIGIN
        1 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
       61 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
      121 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
      181 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
      241 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
      301 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
      361 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
      421 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
      481 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
      541 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagaga
      601 agccacctcg cccaatggta ccaacgatgt tccctccacc aaaggtgttc ttatgtaggc
      661 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
      721 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
      781 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
      841 agtttatcat tatcaatact cgccatttca aagaatacgt aaataattaa tagtagtgat
      901 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
      961 ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt
     1021 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     1081 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     1141 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     1201 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     1261 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     1321 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     1381 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     1441 tttatagtta gtcttttttt tagttttaaa acaccagaac ttagtttcga cggattctag
     1501 aactagtgga tcccccatca caaaatgcag aagcgaatag ccttgtcgtt tccagaagag
     1561 gtactagagc atgtgttctc gtttattcag ctggataagg ataggaactc agtctctctg
     1621 gtgtgcaagt catggtacga gatcgagcgg tggtgcagga ggaaagtctt catcgggaac
     1681 tgctacgccg tgagtccagc gacggtgatt aggaggttcc cgaaagtgag atccgtggag
     1741 cttaaaggaa aacctcactt tgctgacttt aatttggtac ctgacggatg gggaggttac
     1801 gtgtatccat ggattgaggc catgtcttcg tcttacacgt ggcttgaaga gataaggctg
     1861 aagaggatgg tggtcaccga cgattgcttg gagctcatag ccaagtcttt taagaatttt
     1921 aaggttcttg tgctttcttc ctgcgaaggc ttctccaccg atggtctcgc tgctatcgct
     1981 gccacttgca ggaatctgaa agagcttgac ttacgagaga gtgatgttga cgacgttagt
     2041 ggccactggc ttagccattt cccagataca tacacttctt tggtatcact caatatatct
     2101 tgcttagcat ctgaggtcag tttctctgct ctggaaaggc tggtgactag gtgtcccaat
     2161 ctcaagtctc tcaagcttaa ccgagctgtt ccacttgaaa aattggctac tttacttcaa
     2221 agagcacctc aattggagga attgggcact ggtgggtaca ctgcagaagt gcgaccagat
     2281 gtttactctg gtttatctgt agcgctctct gggtgcaagg aattgaggtg cttatctgga
     2341 ttttgggatg ctgttcctgc ctatcttcca gcagtttatt cggtttgcag tcggcttaca
     2401 actttgaatc tgagttatgc aacagtccag agctatgatc ttgtcaagct tctttgtcaa
     2461 tgccctaaac tgcagcgcct ctgggtgctt gactacatcg aggatgctgg tcttgaggtg
     2521 cttgcttcaa cctgcaagga cctacgcgag ctgagagtgt ttccgtccga gccttttgtc
     2581 atggaaccaa atgtggcatt gacggaacag gggcttgtct ccgtctccat gggctgtcca
     2641 aaactcgagt cggttctcta cttctgccgt caaatgacca atgctgcatt gataaccatt
     2701 gctaggaacc gtcccaacat gactcgcttc cgtttgtgca tcattgagcc aaaagcccca
     2761 gactatctga ctctagagcc actggatatt ggatttggag ccatagtaga gcactgcaag
     2821 gatctccgtc gcctctctct atctggcctc ttgaccgaca aggtttttga atacattggg
     2881 acatatgcca agaagatgga aatgctctca gtggcatttg caggagacag tgacttaggc
     2941 atgcatcatg ttttgtccgg gtgcgatagc ttgaggaaac tagagataag ggactgcccg
     3001 tttggagaca aggcgctttt ggccaatgct tcaaagctgg agacaatgcg atccctttgg
     3061 atgtcttctt gttccgtgag ttttggagcc tgcaagttac taggacagaa gatgccaaag
     3121 ctgaatgtgg aagtcatcga tgaacggggt gcaccggact cgagaccaga gagctgccct
     3181 gttgagagag tcttcatata ccgaacagtg gctggtcctc gatttgacat gcctggcttc
     3241 gtctggaaca tggaccaaga ctcaacaatg aggttttcca ggcaaatcat tactactaac
     3301 ggattaggat ctggtggaga ttacaaggat catgatggtg attacaagga tcacgacatc
     3361 gactacaagg atgacgatga caagcaccat catcaccacc attgattcta tgtcaggctc
     3421 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     3481 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     3541 acgtttctcg ttcagctttc ttgtacaaag tggtgatggg ctgcaggaat tcgatttcaa
     3601 gcttatcgat accgtcgacc tcgagtcatg taattagtta tgtcacgctt acattcacgc
     3661 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     3721 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     3781 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     3841 aaggttttgg gacgctcgaa ggctttaatt tgtgacaccg attatttaaa gctgcagcat
     3901 acgatatata tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa
     3961 cagtatgata ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag
     4021 gcgcgctttc cttttttctt tttgcttttt cttttttttt ctcttgaact cgagaaaaaa
     4081 aatataaaag agatggagga acgggaaaaa gttagttgtg gtgataggtg gcaagtggta
     4141 ttccgtaaga acaacaagaa aagcatttca tattatggct gaactgagcg aacaagtgca
     4201 aaatttaagc atcaacgaca acaacgagaa tggttatgtt cctcctcact taagaggaaa
     4261 accaagaagt gccagaaata acagtagcaa ctacaataac aacaacggcg gctacaacgg
     4321 tggccgtggc ggtggcagct tctttagcaa caaccgtcgt ggtggttacg gcaacggtgg
     4381 tttcttcggt ggaaacaacg gtggcagcag atctaacggc cgttctggtg gtagatggat
     4441 cgatggcaaa catgtcccag ctccaagaaa cgaaaaggcc gagatcgcca tatttggtgt
     4501 ccccgaggat ccaaatttcc aatcttctgg tattaacttc gataactacg atgatattcc
     4561 agtggacgcc tctggtaagg atgttcctga accaatcaca gaatttacct cacctccatt
     4621 ggacggattg ttattggaaa acatcaaatt ggcccgtttc accaagccaa cacctgtgca
     4681 aaaatactcc gtccctatcg ttgccaacgg cagagatttg atggcctgtg cgcagaccgg
     4741 ttctggtaag actggtgggt ttttattccc agtgttgtcc gaatcattta agactggacc
     4801 atctcctcaa ccagagtctc aaggctcctt ttaccaaaga aaggcctacc caactgctgt
     4861 cattatggct ccagtttaaa ccatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4921 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4981 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5041 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5101 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5161 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5221 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5701 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5761 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5821 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5881 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5941 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6001 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6061 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6181 taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7021 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7081 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     7141 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     7201 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     7261 catcagagca gattgtactg agagtgcacc ataattcgtt taaaccgttt taagagcttg
     7321 gtgagcgcta ggagtcactg ccaggtatcg tttgaacacg gcattagtca gggaagtcat
     7381 aacacagtcc tttcccgcaa ttttcttttt ctattactct tggcctcctc tagtacactc
     7441 tatatttttt tatgcctcgg taatgatttt catttttttt tttccaccta gcggatgact
     7501 cttttttttt cttagcgatt ggcattatca cataatgaat tatacattat ataaagtaat
     7561 gtgatttctt cgaagaatat actaaaaaat gagcaggcaa gataaacgaa ggcaaagatg
     7621 ac
//
