LOCUS       sequence-210056-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector including N-terminally 3xFLAG-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 4827)
  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 4827)
  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: pAN19-3xFLAG-TIR1-NOSt
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             288..353
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      2150..2402
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2427..2444)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2427..2443)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2436..2458)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2652..2671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2771..2793
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2831..2849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2917..3021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3022..3090,3091..3882)
                     /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(3240..3259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4053..4641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4542..4561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4795..4812
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg aggcgcgccg cggccgcaag
      241 cttgcatgcc tgcaggtcga ctctagagga tcccccgggc cgcaatggac tacaaagacc
      301 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagatgcaga
      361 agcgaatagc cttgtcgttt ccagaagagg tactagagca tgtgttctcg tttattcagc
      421 tggataagga taggaactca gtctctctgg tgtgcaagtc atggtacgag atcgagcggt
      481 ggtgcaggag gaaagtcttc atcgggaact gctacgccgt gagtccagcg acggtgatta
      541 ggaggttccc gaaagtgaga tccgtggagc ttaaaggaaa acctcacttt gctgacttta
      601 atttggtacc tgacggatgg ggaggttacg tgtatccatg gattgaggcc atgtcttcgt
      661 cttacacgtg gcttgaagag ataaggctga agaggatggt ggtcaccgac gattgcttgg
      721 agctcatagc caagtctttt aagaatttta aggtactagt gctttcttcc tgcgaaggct
      781 tctccaccga tggtctcgct gctatcgctg ccacttgcag gaatctgaaa gagcttgact
      841 tacgagagag tgatgttgac gacgttagtg gccactggct tagccatttc ccagatacat
      901 acacttcttt ggtatcactc aatatatctt gcttagcatc tgaggtcagt ttctctgctc
      961 tggaaaggct ggtgactagg tgtcccaatc tcaagtctct caagcttaac cgagctgttc
     1021 cacttgaaaa attggctact ttacttcaaa gagcacctca attggaggaa ttgggcactg
     1081 gtgggtacac tgcagaagtg cgaccagatg tttactctgg tttatctgta gcgctctctg
     1141 ggtgcaagga attgaggtgc ttatctggat tttgggatgc tgttcctgcc tatcttccag
     1201 cagtttattc ggtttgcagt cggcttacaa ctttgaatct gagttatgca acagtccaga
     1261 gctatgatct tgtcaagctt ctttgtcaat gccctaaact gcagcgcctc tgggtgcttg
     1321 actacatcga ggatgctggt cttgaggtgc ttgcttcaac ctgcaaggac ctacgcgagc
     1381 tgagagtgtt tccgtccgag ccttttgtca tggaaccaaa tgtggcattg acggaacagg
     1441 ggcttgtctc cgtctccatg ggctgtccaa aactcgagtc ggttctctac ttctgccgtc
     1501 aaatgaccaa tgctgcattg ataaccattg ctaggaaccg tcccaacatg actcgcttcc
     1561 gtttgtgcat cattgagcca aaagccccag actatctgac tctagagcca ctggatattg
     1621 gatttggagc catagtagag cactgcaagg atctccgtcg cctctctcta tctggcctct
     1681 tgaccgacaa ggtttttgaa tacattggga catatgccaa gaagatggaa atgctctcag
     1741 tggcatttgc aggagacagt gacttaggca tgcatcatgt tttgtccggg tgcgatagct
     1801 tgaggaaact agagataagg gactgcccgt ttggagacaa ggcgcttttg gccaatgctt
     1861 caaagctgga gacaatgcga tccctttgga tgtcttcttg ttccgtgagt tttggagcct
     1921 gcaagttact aggacagaag atgccaaagc tgaatgtgga agtcatcgat gaacggggtg
     1981 caccggactc gagaccagag agctgccctg ttgagagagt cttcatatac cgaacagtgg
     2041 ctggtcctcg atttgacatg cctggcttcg tctggaacat ggaccaagac tcaacaatga
     2101 ggttttccag gcaaatcatt actactaacg gattataaga gctctccccg atcgttcaaa
     2161 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     2221 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     2281 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     2341 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     2401 tcgaattcgg cgcgccgcgg ccgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2461 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2521 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2581 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     2641 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2701 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2761 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2821 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2881 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3001 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3061 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3121 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3181 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3241 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3301 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3361 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3421 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3481 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3541 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3601 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3661 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3721 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3781 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3841 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3901 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3961 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4801 tcagtgagcg aggaagcgga agagcgc
//
