LOCUS       sequence-210051-        6591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector including C-terminally GUS-tagged Arabidopsis auxin 
            receptor TIR1 with the F79G mutation [ccvTIR1] 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-ccvTIR1-GUS-NOSt
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2094..3905
                     /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      3917..4169
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4194..4211)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4194..4210)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4203..4225)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4419..4438)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4538..4560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4598..4616)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4684..4788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4789..4857,4858..5649)
                     /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(5007..5026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5820..6408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6309..6328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6562..6579
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgaggcgcg ccgcggccgc
      241 aagcttgcat gcctgcaggt cgactctaga ggatcccccg ggccgcaatg cagaagcgaa
      301 tagccttgtc gtttccagaa gaggtactag agcatgtgtt ctcgtttatt cagctggata
      361 aggataggaa ctcagtctct ctggtgtgca agtcatggta cgagatcgag cggtggtgca
      421 ggaggaaagt cttcatcggg aactgctacg ccgtgagtcc agcgacggtg attaggaggt
      481 tcccgaaagt gagatccgtg gagcttaaag gaaaacctca cggtgctgac tttaatttgg
      541 tacctgacgg atggggaggt tacgtgtatc catggattga ggccatgtct tcgtcttaca
      601 cgtggcttga agagataagg ctgaagagga tggtggtcac cgacgattgc ttggagctca
      661 tagccaagtc ttttaagaat tttaaggtac tagtgctttc ttcctgcgaa ggcttctcca
      721 ccgatggtct cgctgctatc gctgccactt gcaggaatct gaaagagctt gacttacgag
      781 agagtgatgt tgacgacgtt agtggccact ggcttagcca tttcccagat acatacactt
      841 ctttggtatc actcaatata tcttgcttag catctgaggt cagtttctct gctctggaaa
      901 ggctggtgac taggtgtccc aatctcaagt ctctcaagct taaccgagct gttccacttg
      961 aaaaattggc tactttactt caaagagcac ctcaattgga ggaattgggc actggtgggt
     1021 acactgcaga agtgcgacca gatgtttact ctggtttatc tgtagcgctc tctgggtgca
     1081 aggaattgag gtgcttatct ggattttggg atgctgttcc tgcctatctt ccagcagttt
     1141 attcggtttg cagtcggctt acaactttga atctgagtta tgcaacagtc cagagctatg
     1201 atcttgtcaa gcttctttgt caatgcccta aactgcagcg cctctgggtg cttgactaca
     1261 tcgaggatgc tggtcttgag gtgcttgctt caacctgcaa ggacctacgc gagctgagag
     1321 tgtttccgtc cgagcctttt gtcatggaac caaatgtggc attgacggaa caggggcttg
     1381 tctccgtctc catgggctgt ccaaaactcg agtcggttct ctacttctgc cgtcaaatga
     1441 ccaatgctgc attgataacc attgctagga accgtcccaa catgactcgc ttccgtttgt
     1501 gcatcattga gccaaaagcc ccagactatc tgactctaga gccactggat attggatttg
     1561 gagccatagt agagcactgc aaggatctcc gtcgcctctc tctatctggc ctcttgaccg
     1621 acaaggtttt tgaatacatt gggacatatg ccaagaagat ggaaatgctc tcagtggcat
     1681 ttgcaggaga cagtgactta ggcatgcatc atgttttgtc cgggtgcgat agcttgagga
     1741 aactagagat aagggactgc ccgtttggag acaaggcgct tttggccaat gcttcaaagc
     1801 tggagacaat gcgatccctt tggatgtctt cttgttccgt gagttttgga gcctgcaagt
     1861 tactaggaca gaagatgcca aagctgaatg tggaagtcat cgatgaacgg ggtgcaccgg
     1921 actcgagacc agagagctgc cctgttgaga gagtcttcat ataccgaaca gtggctggtc
     1981 ctcgatttga catgcctggc ttcgtctgga acatggacca agactcaaca atgaggtttt
     2041 ccaggcaaat cattactact aacggattag tcgagggtgg ctcgaccggt ggtatgttac
     2101 gtcctgtaga aaccccaacc cgtgaaatca aaaaactcga cggcctgtgg gcattcagtc
     2161 tggatcgcga aaactgtgga attgatcagc gttggtggga aagcgcgtta caagaaagcc
     2221 gggcaattgc tgtgccaggc agttttaacg atcagttcgc cgatgcagat attcgtaatt
     2281 atgcgggcaa cgtctggtat cagcgcgaag tctttatacc gaaaggttgg gcaggccagc
     2341 gtatcgtgct gcgtttcgat gcggtcactc attacggcaa agtgtgggtc aataatcagg
     2401 aagtgatgga gcatcagggc ggctatacgc catttgaagc cgatgtcacg ccgtatgtta
     2461 ttgccgggaa aagtgtacgt atcaccgttt gtgtgaacaa cgaactgaac tggcagacta
     2521 tcccgccggg aatggtgatt accgacgaaa acggcaagaa aaagcagtct tacttccatg
     2581 atttctttaa ctatgccgga atccatcgca gcgtaatgct ctacaccacg ccgaacacct
     2641 gggtggacga tatcaccgtg gtgacgcatg tcgcgcaaga ctgtaaccac gcgtctgttg
     2701 actggcaggt ggtggccaat ggtgatgtca gcgttgaact gcgtgatgcg gatcaacagg
     2761 tggttgcaac tggacaaggc actagcggga ctttgcaagt ggtgaatccg cacctctggc
     2821 aaccgggtga aggttatctc tatgaactgt gcgtcacagc caaaagccag acagagtgtg
     2881 atatctaccc gcttcgcgtc ggcatccggt cagtggcagt gaagggcgaa cagttcctga
     2941 ttaaccacaa accgttctac tttactggct ttggtcgtca tgaagatgcg gacttgcgtg
     3001 gcaaaggatt cgataacgtg ctgatggtgc acgaccacgc attaatggac tggattgggg
     3061 ccaactccta ccgtacctcg cattaccctt acgctgaaga gatgctcgac tgggcagatg
     3121 aacatggcat cgtggtgatt gatgaaactg ctgctgtcgg ctttaacctc tctttaggca
     3181 ttggtttcga agcgggcaac aagccgaaag aactgtacag cgaagaggca gtcaacgggg
     3241 aaactcagca agcgcactta caggcgatta aagagctgat agcgcgtgac aaaaaccacc
     3301 caagcgtggt gatgtggagt attgccaacg aaccggatac ccgtccgcaa ggtgcacggg
     3361 aatatttcgc gccactggcg gaagcaacgc gtaaactcga cccgacgcgt ccgatcacct
     3421 gcgtcaatgt aatgttctgc gacgctcaca ccgataccat cagcgatctc tttgatgtgc
     3481 tgtgcctgaa ccgttattac ggatggtatg tccaaagcgg cgatttggaa acggcagaga
     3541 aggtactgga aaaagaactt ctggcctggc aggagaaact gcatcagccg attatcatca
     3601 ccgaatacgg cgtggatacg ttagccgggc tgcactcaat gtacaccgac atgtggagtg
     3661 aagagtatca gtgtgcatgg ctggatatgt atcaccgcgt ctttgatcgc gtcagcgccg
     3721 tcgtcggtga acaggtatgg aatttcgccg attttgcgac ctcgcaaggc atattgcgcg
     3781 ttggcggtaa caagaaaggg atcttcactc gcgaccgcaa accgaagtcg gcggcttttc
     3841 tgctgcaaaa acgctggact ggcatgaact tcggtgaaaa accgcagcag ggaggcaaac
     3901 aatgagagct ctccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     3961 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     4021 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     4081 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     4141 gcgcgcggtg tcatctatgt tactagatcg aattcggcgc gccgcggccg ctcactggcc
     4201 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4261 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4321 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     4381 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     4441 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4501 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4561 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4621 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4681 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4741 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4801 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4861 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4921 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4981 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5041 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5101 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5161 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5221 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5281 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5341 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5401 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5461 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5521 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5581 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5641 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5701 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5761 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5821 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5881 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5941 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6001 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6061 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6121 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6181 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6241 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6301 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6361 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6421 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6481 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6541 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga g
//
