LOCUS       sequence-210059-        6591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector including C-terminally GUS-tagged Arabidopsis auxin 
            receptor TIR1 [Wild type] and NOS terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6591)
  AUTHORS   Uchida N, Takahashi K, Iwasaki R, Yamada R, Yoshimura M, Endo TA, 
            Kimura S, Zhang H, Nomoto M, Tada Y, Kinoshita T, Itami K, Hagihara 
            S, Torii KU
  TITLE     Chemical hijacking of auxin signaling with an engineered auxin-TIR1 
            pair.
  JOURNAL   Nat Chem Biol. 2018 Mar;14(3):299-305. doi: 10.1038/nchembio.2555. 
            Epub 2018 Jan 22.
  PUBMED    29355850
REFERENCE   2  (bases 1 to 6591)
  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: 
            "10.1038/nchembio.2555"; journalName: "Nat Chem Biol"; date: 
            "2018-03"; volume: "14"; issue: "3"; pages: "299-305"
COMMENT     Sequence Label: pAN19-TIR1-GUS-NOSt
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..45
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    162..183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(198..203,204..221,222..228)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    236..252
                     /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)."
     primer_bind     241..263
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     260..276
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     260..276
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2151..3962
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     terminator      3974..4226
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4251..4268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4251..4267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4260..4282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4476..4495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4595..4617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4655..4673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4741..4845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4846..4914,4915..5706)
                     /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     complement(5064..5083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5877..6465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6366..6385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
       61 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      121 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      181 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      241 agcggataac aatttcacac aggaaacagc tatgaccatg aggcgcgccg cggccgcaag
      301 cttgcatgcc tgcaggtcga ctctagagga tcccccgggc cgcaatgcag aagcgaatag
      361 ccttgtcgtt tccagaagag gtactagagc atgtgttctc gtttattcag ctggataagg
      421 ataggaactc agtctctctg gtgtgcaagt catggtacga gatcgagcgg tggtgcagga
      481 ggaaagtctt catcgggaac tgctacgccg tgagtccagc gacggtgatt aggaggttcc
      541 cgaaagtgag atccgtggag cttaaaggaa aacctcactt tgctgacttt aatttggtac
      601 ctgacggatg gggaggttac gtgtatccat ggattgaggc catgtcttcg tcttacacgt
      661 ggcttgaaga gataaggctg aagaggatgg tggtcaccga cgattgcttg gagctcatag
      721 ccaagtcttt taagaatttt aaggtactag tgctttcttc ctgcgaaggc ttctccaccg
      781 atggtctcgc tgctatcgct gccacttgca ggaatctgaa agagcttgac ttacgagaga
      841 gtgatgttga cgacgttagt ggccactggc ttagccattt cccagataca tacacttctt
      901 tggtatcact caatatatct tgcttagcat ctgaggtcag tttctctgct ctggaaaggc
      961 tggtgactag gtgtcccaat ctcaagtctc tcaagcttaa ccgagctgtt ccacttgaaa
     1021 aattggctac tttacttcaa agagcacctc aattggagga attgggcact ggtgggtaca
     1081 ctgcagaagt gcgaccagat gtttactctg gtttatctgt agcgctctct gggtgcaagg
     1141 aattgaggtg cttatctgga ttttgggatg ctgttcctgc ctatcttcca gcagtttatt
     1201 cggtttgcag tcggcttaca actttgaatc tgagttatgc aacagtccag agctatgatc
     1261 ttgtcaagct tctttgtcaa tgccctaaac tgcagcgcct ctgggtgctt gactacatcg
     1321 aggatgctgg tcttgaggtg cttgcttcaa cctgcaagga cctacgcgag ctgagagtgt
     1381 ttccgtccga gccttttgtc atggaaccaa atgtggcatt gacggaacag gggcttgtct
     1441 ccgtctccat gggctgtcca aaactcgagt cggttctcta cttctgccgt caaatgacca
     1501 atgctgcatt gataaccatt gctaggaacc gtcccaacat gactcgcttc cgtttgtgca
     1561 tcattgagcc aaaagcccca gactatctga ctctagagcc actggatatt ggatttggag
     1621 ccatagtaga gcactgcaag gatctccgtc gcctctctct atctggcctc ttgaccgaca
     1681 aggtttttga atacattggg acatatgcca agaagatgga aatgctctca gtggcatttg
     1741 caggagacag tgacttaggc atgcatcatg ttttgtccgg gtgcgatagc ttgaggaaac
     1801 tagagataag ggactgcccg tttggagaca aggcgctttt ggccaatgct tcaaagctgg
     1861 agacaatgcg atccctttgg atgtcttctt gttccgtgag ttttggagcc tgcaagttac
     1921 taggacagaa gatgccaaag ctgaatgtgg aagtcatcga tgaacggggt gcaccggact
     1981 cgagaccaga gagctgccct gttgagagag tcttcatata ccgaacagtg gctggtcctc
     2041 gatttgacat gcctggcttc gtctggaaca tggaccaaga ctcaacaatg aggttttcca
     2101 ggcaaatcat tactactaac ggattagtcg agggtggctc gaccggtggt atgttacgtc
     2161 ctgtagaaac cccaacccgt gaaatcaaaa aactcgacgg cctgtgggca ttcagtctgg
     2221 atcgcgaaaa ctgtggaatt gatcagcgtt ggtgggaaag cgcgttacaa gaaagccggg
     2281 caattgctgt gccaggcagt tttaacgatc agttcgccga tgcagatatt cgtaattatg
     2341 cgggcaacgt ctggtatcag cgcgaagtct ttataccgaa aggttgggca ggccagcgta
     2401 tcgtgctgcg tttcgatgcg gtcactcatt acggcaaagt gtgggtcaat aatcaggaag
     2461 tgatggagca tcagggcggc tatacgccat ttgaagccga tgtcacgccg tatgttattg
     2521 ccgggaaaag tgtacgtatc accgtttgtg tgaacaacga actgaactgg cagactatcc
     2581 cgccgggaat ggtgattacc gacgaaaacg gcaagaaaaa gcagtcttac ttccatgatt
     2641 tctttaacta tgccggaatc catcgcagcg taatgctcta caccacgccg aacacctggg
     2701 tggacgatat caccgtggtg acgcatgtcg cgcaagactg taaccacgcg tctgttgact
     2761 ggcaggtggt ggccaatggt gatgtcagcg ttgaactgcg tgatgcggat caacaggtgg
     2821 ttgcaactgg acaaggcact agcgggactt tgcaagtggt gaatccgcac ctctggcaac
     2881 cgggtgaagg ttatctctat gaactgtgcg tcacagccaa aagccagaca gagtgtgata
     2941 tctacccgct tcgcgtcggc atccggtcag tggcagtgaa gggcgaacag ttcctgatta
     3001 accacaaacc gttctacttt actggctttg gtcgtcatga agatgcggac ttgcgtggca
     3061 aaggattcga taacgtgctg atggtgcacg accacgcatt aatggactgg attggggcca
     3121 actcctaccg tacctcgcat tacccttacg ctgaagagat gctcgactgg gcagatgaac
     3181 atggcatcgt ggtgattgat gaaactgctg ctgtcggctt taacctctct ttaggcattg
     3241 gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga agaggcagtc aacggggaaa
     3301 ctcagcaagc gcacttacag gcgattaaag agctgatagc gcgtgacaaa aaccacccaa
     3361 gcgtggtgat gtggagtatt gccaacgaac cggatacccg tccgcaaggt gcacgggaat
     3421 atttcgcgcc actggcggaa gcaacgcgta aactcgaccc gacgcgtccg atcacctgcg
     3481 tcaatgtaat gttctgcgac gctcacaccg ataccatcag cgatctcttt gatgtgctgt
     3541 gcctgaaccg ttattacgga tggtatgtcc aaagcggcga tttggaaacg gcagagaagg
     3601 tactggaaaa agaacttctg gcctggcagg agaaactgca tcagccgatt atcatcaccg
     3661 aatacggcgt ggatacgtta gccgggctgc actcaatgta caccgacatg tggagtgaag
     3721 agtatcagtg tgcatggctg gatatgtatc accgcgtctt tgatcgcgtc agcgccgtcg
     3781 tcggtgaaca ggtatggaat ttcgccgatt ttgcgacctc gcaaggcata ttgcgcgttg
     3841 gcggtaacaa gaaagggatc ttcactcgcg accgcaaacc gaagtcggcg gcttttctgc
     3901 tgcaaaaacg ctggactggc atgaacttcg gtgaaaaacc gcagcaggga ggcaaacaat
     3961 gagagctctc cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     4021 tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat
     4081 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     4141 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     4201 cgcggtgtca tctatgttac tagatcgaat tcggcgcgcc gcggccgctc actggccgtc
     4261 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4321 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4381 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     4441 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4501 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4561 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4621 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4681 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4741 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4801 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4861 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4921 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4981 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5041 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5101 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5161 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5221 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5281 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5341 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5401 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5461 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5521 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5581 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5641 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5701 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5761 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5821 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5881 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5941 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6001 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6061 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6121 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6181 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6241 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6301 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6361 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6421 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6481 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6541 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac c
//
