LOCUS       sequence-210054-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector including N-terminally 3xFLAG-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 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-ccvTIR1-NOSt
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..20
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    137..158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(173..178,179..196,197..203)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    211..227
                     /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     216..238
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     235..251
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     235..251
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             323..388
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      2185..2437
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2462..2479)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2462..2478)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2471..2493)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2687..2706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2806..2828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2866..2884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2952..3056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3057..3125,3126..3917)
                     /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(3275..3294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4088..4676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4577..4596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
       61 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
      121 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
      181 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      241 acagctatga ccatgaggcg cgccgcggcc gcaagcttgc atgcctgcag gtcgactcta
      301 gaggatcccc cgggccgcaa tggactacaa agaccatgac ggtgattata aagatcatga
      361 catcgattac aaggatgacg atgacaagat gcagaagcga atagccttgt cgtttccaga
      421 agaggtacta gagcatgtgt tctcgtttat tcagctggat aaggatagga actcagtctc
      481 tctggtgtgc aagtcatggt acgagatcga gcggtggtgc aggaggaaag tcttcatcgg
      541 gaactgctac gccgtgagtc cagcgacggt gattaggagg ttcccgaaag tgagatccgt
      601 ggagcttaaa ggaaaacctc acggtgctga ctttaatttg gtacctgacg gatggggagg
      661 ttacgtgtat ccatggattg aggccatgtc ttcgtcttac acgtggcttg aagagataag
      721 gctgaagagg atggtggtca ccgacgattg cttggagctc atagccaagt cttttaagaa
      781 ttttaaggta ctagtgcttt cttcctgcga aggcttctcc accgatggtc tcgctgctat
      841 cgctgccact tgcaggaatc tgaaagagct tgacttacga gagagtgatg ttgacgacgt
      901 tagtggccac tggcttagcc atttcccaga tacatacact tctttggtat cactcaatat
      961 atcttgctta gcatctgagg tcagtttctc tgctctggaa aggctggtga ctaggtgtcc
     1021 caatctcaag tctctcaagc ttaaccgagc tgttccactt gaaaaattgg ctactttact
     1081 tcaaagagca cctcaattgg aggaattggg cactggtggg tacactgcag aagtgcgacc
     1141 agatgtttac tctggtttat ctgtagcgct ctctgggtgc aaggaattga ggtgcttatc
     1201 tggattttgg gatgctgttc ctgcctatct tccagcagtt tattcggttt gcagtcggct
     1261 tacaactttg aatctgagtt atgcaacagt ccagagctat gatcttgtca agcttctttg
     1321 tcaatgccct aaactgcagc gcctctgggt gcttgactac atcgaggatg ctggtcttga
     1381 ggtgcttgct tcaacctgca aggacctacg cgagctgaga gtgtttccgt ccgagccttt
     1441 tgtcatggaa ccaaatgtgg cattgacgga acaggggctt gtctccgtct ccatgggctg
     1501 tccaaaactc gagtcggttc tctacttctg ccgtcaaatg accaatgctg cattgataac
     1561 cattgctagg aaccgtccca acatgactcg cttccgtttg tgcatcattg agccaaaagc
     1621 cccagactat ctgactctag agccactgga tattggattt ggagccatag tagagcactg
     1681 caaggatctc cgtcgcctct ctctatctgg cctcttgacc gacaaggttt ttgaatacat
     1741 tgggacatat gccaagaaga tggaaatgct ctcagtggca tttgcaggag acagtgactt
     1801 aggcatgcat catgttttgt ccgggtgcga tagcttgagg aaactagaga taagggactg
     1861 cccgtttgga gacaaggcgc ttttggccaa tgcttcaaag ctggagacaa tgcgatccct
     1921 ttggatgtct tcttgttccg tgagttttgg agcctgcaag ttactaggac agaagatgcc
     1981 aaagctgaat gtggaagtca tcgatgaacg gggtgcaccg gactcgagac cagagagctg
     2041 ccctgttgag agagtcttca tataccgaac agtggctggt cctcgatttg acatgcctgg
     2101 cttcgtctgg aacatggacc aagactcaac aatgaggttt tccaggcaaa tcattactac
     2161 taacggatta taagagctct ccccgatcgt tcaaacattt ggcaataaag tttcttaaga
     2221 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag
     2281 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga
     2341 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat
     2401 aaattatcgc gcgcggtgtc atctatgtta ctagatcgaa ttcggcgcgc cgcggccgct
     2461 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2521 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2581 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2641 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     2701 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2761 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2821 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     2881 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2941 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3001 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3061 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3121 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3181 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3241 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3301 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3361 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3421 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3481 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3541 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3601 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3661 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3721 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3781 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3841 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3901 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3961 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4021 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4081 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4141 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4201 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4261 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4321 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4381 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4441 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4501 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4561 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4621 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4681 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4741 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4801 ccgctcgccg cagccgaacg accgagc
//
