LOCUS       sequence-370254-        7559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zea Mays Nuclear Auxin Signaling Receptor AFB2/3 A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  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 7559)
  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-ZmAFB2/3 a
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      60..225
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(115..139)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        248..891
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     785..808
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     897..913
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     897..913
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2657..2722
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2723..2740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2788..2912)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2945..2961)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2946..2962)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2964..3210
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2980..2998)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4224..4240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4224..4240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4237..4259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4248..4264
                     /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(4272..4278,4279..4296,4297..4302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4317..4338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4455..4472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4626..5214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4706..4725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5385..6176,6177..6245))
                     /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     6008..6027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6246..6350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6418..6436
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6474..6496)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6596..6615
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6768..6955
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(6956..7559,1..59)
                     /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 aagccacctc gcccaatggt accaacgatg ttccctccac caaaggtgtt cttatgtagg
       61 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      121 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      181 tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct
      241 cagtttatca ttatcaatac tcgccatttc aaagaatacg taaataatta atagtagtga
      301 ttttcctaac tttatttagt caaaaaatta gccttttaat tctgctgtaa cccgtacatg
      361 cccaaaatag ggggcgggtt acacagaata tataacatcg taggtgtctg ggtgaacagt
      421 ttattcctgg catccactaa atataatgga gcccgctttt taagctggca tccagaaaaa
      481 aaaagaatcc cagcaccaaa atattgtttt cttcaccaac catcagttca taggtccatt
      541 ctcttagcgc aactacagag aacaggggca caaacaggca aaaaacgggc acaacctcaa
      601 tggagtgatg caacctgcct ggagtaaatg atgacacaag gcaattgacc cacgcatgta
      661 tctatctcat tttcttacac cttctattac cttctgctct ctctgatttg gaaaaagctg
      721 aaaaaaaagg ttgaaaccag ttccctgaaa ttattcccct acttgactaa taagtatata
      781 aagacggtag gtattgattg taattctgta aatctatttc ttaaacttct taaattctac
      841 ttttatagtt agtctttttt ttagttttaa aacaccagaa cttagtttcg acggattcta
      901 gaactagtgg atcccccatc acaaaatggc gtactttcca gacgaagtag ttgggtacat
      961 tttagggtac gtaacatctc accaagatag aaatgcggtt agcttggttt gcagggcatg
     1021 gtacgacatt gagagacacg ggaggcatag tgtactagtt aggaattgct atgcagtttg
     1081 tcctgaacgt gttcacatga ggtttcctaa catgagggca ctgtcactaa agggtaagcc
     1141 ccattttgca gaattcaacc ttgtacccgc cggctgggga gctaccgcca acccgtgggt
     1201 tgatgcctgt gctagagctt gtccaggact ggaggaactg aggttgaaat ttatggtagt
     1261 gactgacgag tgcttaaaat tattatccct ttccttcacg aattttaaga gcttggttct
     1321 agtatgttgt gaggggtttt ccacgactgg tttagcaaat atagctacaa actgtagatt
     1381 ccttaaggag ctagatctgc agaaatcttg tgtcaagcac caagatcacc agtggattaa
     1441 ctgctttcct aaatcatcta catccctaga gtgcctaaat ttcagttgcc ttaccggaga
     1501 ggtgaatgcc gtggcgctag aggaattggt cgcgcgttca ccaaacttaa aatccttgag
     1561 actaaatctg gctgtcccgt tcgacgtcct aagccgtata ttgtctcgta cgcccaaact
     1621 ggaagattta ggtactggtt cttttttgca gggaaatgac ccagcagcct atgcatccct
     1681 gtgccgtgcg cttgaaaact gtactagcct taaaagcatt tcagggtttt gggacgctcc
     1741 agggttctac gtgcaaggga tactaagtaa ttgcaaaatc aggaatctaa cctgccttaa
     1801 tttgagctat gcaactttaa tccaatcaac acaactaata ggaatcataa gacactgcaa
     1861 gaaactacac gtcttgtggg tgctagacca tataggtgat gagggtctga aggccgtgag
     1921 cttctcttgt cctgatctgc aggagcttcg tgtctacccg tctgtggtcg cacccagagg
     1981 gacagtaacc gaagaggggc tagtggcatt gagctcctgt cgtaagctac aacatgtgtt
     2041 gttttcctgc gttcgtatga ctaacacggc cctaatgaca atcgcaagat actgtcccag
     2101 gctaacttct ttccgtctgt gtattagaaa accaagaagt gcggacgcgg taacaggtca
     2161 gccccttgac gagggattcg gtgcaattgt ccgttcttgc agggggttga gaagattggc
     2221 gatgagcggc ctactaacag attccgtttt cctttatatt ggtatgtacg ctgaaaaatt
     2281 ggagatgttg agcgtcactt tcgcaggtga caccgatgat ggaatggtgt atgtcttaaa
     2341 cggatgcaga aaccttaaga agctagtcat taaggaatca ccattcggcg acgccgcgtt
     2401 actggctgga gcccacagat acgagagcat gaggagtctg tggatgtcct cttgtcaaat
     2461 aactttgggg ggttgcaagg ctttagccgc cacgatgccc aacattaatg ttgaagtgat
     2521 aggtggagcc tcttttggag ctatggatgg aggagtaagt ggagctcgta aagttgatat
     2581 gttatatttg tatagaactt tggcggggcc tagatgcgat acccctgggt tcgtgagtat
     2641 tctaggatct ggtggagatt acaaggatca tgatggtgat tacaaggatc acgacatcga
     2701 ctacaaggat gacgatgaca agcaccatca tcaccaccat tgattctatg tcaggctccc
     2761 ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat
     2821 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
     2881 gtttctcgtt cagctttctt gtacaaagtg gtgatgggct gcaggaattc gatttcaagc
     2941 ttatcgatac cgtcgacctc gagtcatgta attagttatg tcacgcttac attcacgccc
     3001 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     3061 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     3121 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     3181 ggttttggga cgctcgaagg ctttaatttg tgacaccgat tatttaaagc tgcagcatac
     3241 gatatatata catgtgtata tatgtatacc tatgaatgtc agtaagtatg tatacgaaca
     3301 gtatgatact gaagatgaca aggtaatgca tcattctata cgtgtcattc tgaacgaggc
     3361 gcgctttcct tttttctttt tgctttttct ttttttttct cttgaactcg agaaaaaaaa
     3421 tataaaagag atggaggaac gggaaaaagt tagttgtggt gataggtggc aagtggtatt
     3481 ccgtaagaac aacaagaaaa gcatttcata ttatggctga actgagcgaa caagtgcaaa
     3541 atttaagcat caacgacaac aacgagaatg gttatgttcc tcctcactta agaggaaaac
     3601 caagaagtgc cagaaataac agtagcaact acaataacaa caacggcggc tacaacggtg
     3661 gccgtggcgg tggcagcttc tttagcaaca accgtcgtgg tggttacggc aacggtggtt
     3721 tcttcggtgg aaacaacggt ggcagcagat ctaacggccg ttctggtggt agatggatcg
     3781 atggcaaaca tgtcccagct ccaagaaacg aaaaggccga gatcgccata tttggtgtcc
     3841 ccgaggatcc aaatttccaa tcttctggta ttaacttcga taactacgat gatattccag
     3901 tggacgcctc tggtaaggat gttcctgaac caatcacaga atttacctca cctccattgg
     3961 acggattgtt attggaaaac atcaaattgg cccgtttcac caagccaaca cctgtgcaaa
     4021 aatactccgt ccctatcgtt gccaacggca gagatttgat ggcctgtgcg cagaccggtt
     4081 ctggtaagac tggtgggttt ttattcccag tgttgtccga atcatttaag actggaccat
     4141 ctcctcaacc agagtctcaa ggctcctttt accaaagaaa ggcctaccca actgctgtca
     4201 ttatggctcc agtttaaacc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4261 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4321 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4381 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4441 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4501 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4561 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4621 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4681 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4741 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4801 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4861 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4921 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4981 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5041 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5101 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5161 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5221 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5281 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5341 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5401 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5461 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5521 ccgcgagatc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5581 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5641 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5701 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5761 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5821 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5881 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5941 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6001 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6061 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6121 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6181 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6241 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6301 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6361 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6421 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6481 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6541 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6601 tcagagcaga ttgtactgag agtgcaccat aattcgttta aaccgtttta agagcttggt
     6661 gagcgctagg agtcactgcc aggtatcgtt tgaacacggc attagtcagg gaagtcataa
     6721 cacagtcctt tcccgcaatt ttctttttct attactcttg gcctcctcta gtacactcta
     6781 tattttttta tgcctcggta atgattttca tttttttttt tccacctagc ggatgactct
     6841 ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat aaagtaatgt
     6901 gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg caaagatgac
     6961 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
     7021 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
     7081 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
     7141 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
     7201 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
     7261 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
     7321 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
     7381 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
     7441 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
     7501 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagag
//
