LOCUS       sequence-370257-        7559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zea Mays Nuclear Auxin Signaling Receptor AFB2/3 b2.
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 b2
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      659..824
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(714..738)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        847..1490
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1384..1407
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1496..1512
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1496..1512
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3256..3321
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3322..3339
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3387..3511)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3544..3560)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3545..3561)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3563..3809
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3579..3597)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4823..4839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4823..4839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4836..4858)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4847..4863
                     /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(4871..4877,4878..4895,4896..4901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4916..4937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5054..5071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5225..5813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5305..5324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5984..6775,6776..6844))
                     /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     6607..6626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6845..6949)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7017..7035
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7073..7095)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7195..7214
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7367..7554
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(7555..7559,1..658)
                     /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 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
       61 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
      121 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
      181 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
      241 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
      301 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
      361 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
      421 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
      481 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
      541 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagaga
      601 agccacctcg cccaatggta ccaacgatgt tccctccacc aaaggtgttc ttatgtaggc
      661 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
      721 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
      781 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
      841 agtttatcat tatcaatact cgccatttca aagaatacgt aaataattaa tagtagtgat
      901 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
      961 ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt
     1021 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     1081 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     1141 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     1201 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     1261 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     1321 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     1381 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     1441 tttatagtta gtcttttttt tagttttaaa acaccagaac ttagtttcga cggattctag
     1501 aactagtgga tcccccatca caaaatgaca tacttcccgg gcgaggtcgt agagcacatc
     1561 ttttcatttt tgccgagtca cagcgacagg aacacggtgt cactagtttg taaactttgg
     1621 tatgacgtgg aaaggctgag tcgtagaggt gtcttcgtag ggaactgtta cgccgtatta
     1681 cctgaaagag tggtgctgcg tttcccaaac gtgaaagctc taacagttaa aggtaagccg
     1741 cactttgccg actttaattt agttccccca gattggggcg gatacgctgg cccatggata
     1801 gaagctacag ccagaagctg tttgtcccta gaggagctga ggatgaaaag gatggtaatc
     1861 agtgacgaga atttggaact tctggcgaga acctttccga aattcaaggt attggtatta
     1921 atatcttgtg aaggtttcag cactgacgga ctagcggcaa tagcgaatca ttgtaaactg
     1981 ttgcgtgaat tagacctaca ggagaacgac gtagaggata ggggtcccag atggctgtcc
     2041 tttcccgact cctgcacctc tttagtttca ctaaatttcg cctgtatcaa gggtgaagtg
     2101 aactctggtg ctctagagcg tctggtcgct aaatcaccta acctgaggag tcttaggtta
     2161 aatcgttccg tctcagtcga caccttgtca aagattcttg aacgtacgcc taacctggag
     2221 gacttaggga ctggtaatct tacggacgag tttcaagccg agagtttcgt acgtcttact
     2281 agcgctctgg agaagtgcaa gaggctgcgt aatcttagcg gcttttggga cgcctcacca
     2341 atctttgtgc cttttatata cccgttatgc catcagttaa cggggcttaa tctgtcctat
     2401 acccctacgc ttgactattc agacttaacc aaaatgataa gtaggtgcgt caagcttcag
     2461 agactatggg ttctagattg cattagtgac aaggggttgc aggtcgtcgc ttcttcctgc
     2521 aaagaccttc aggagttaag agtatttcca tcagagttta acgttgctgg tgcgtttacc
     2581 gtgacagaag aaggtttagt ggcaatttct agcggttgtc ccaagctaag ctcattattg
     2641 tacttttgcc accagatgac caatgaggct ttgacaacgg ttgctaagaa ctgtccctcc
     2701 tttatcagat ttagactgtg catactggag cctaagaagc cagacgccat gacaggccaa
     2761 ccgctagatg aagggtttgg agctatagta cgtgattgca aggggttgcg taggctaagt
     2821 atgagcggcc tacttacaga ccgtgtcttt atgtatataa gaatgtacgc aaaatatctt
     2881 gagatgttgt ctatcgcgtt tgccggagat ggggataaag gtatgatgga cgtgatgaac
     2941 ggctgtaaga atttacgtaa gttagaaatc agggacagcc cgttcggtga ctttgcgctg
     3001 ttgggtaacg tggccaaata tgataccatg cgttccctat ggatgtcttc atgcaacgtg
     3061 acgctgaagg gatgccaagt tttagcttcc aagatgccaa tgcttaacgt ggagataatg
     3121 aacgaacttg atgggtcttc agaaatggag aaccatggca acctgtcaaa agttgataaa
     3181 ctgtacgtct atagaacaac tgcgggtgta agggacgatg cccctaactt tgtacagata
     3241 ctgggatctg gtggagatta caaggatcat gatggtgatt acaaggatca cgacatcgac
     3301 tacaaggatg acgatgacaa gcaccatcat caccaccatt gattctatgt caggctccct
     3361 tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt
     3421 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     3481 tttctcgttc agctttcttg tacaaagtgg tgatgggctg caggaattcg atttcaagct
     3541 tatcgatacc gtcgacctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     3601 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     3661 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     3721 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     3781 gttttgggac gctcgaaggc tttaatttgt gacaccgatt atttaaagct gcagcatacg
     3841 atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt atacgaacag
     3901 tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct gaacgaggcg
     3961 cgctttcctt ttttcttttt gctttttctt tttttttctc ttgaactcga gaaaaaaaat
     4021 ataaaagaga tggaggaacg ggaaaaagtt agttgtggtg ataggtggca agtggtattc
     4081 cgtaagaaca acaagaaaag catttcatat tatggctgaa ctgagcgaac aagtgcaaaa
     4141 tttaagcatc aacgacaaca acgagaatgg ttatgttcct cctcacttaa gaggaaaacc
     4201 aagaagtgcc agaaataaca gtagcaacta caataacaac aacggcggct acaacggtgg
     4261 ccgtggcggt ggcagcttct ttagcaacaa ccgtcgtggt ggttacggca acggtggttt
     4321 cttcggtgga aacaacggtg gcagcagatc taacggccgt tctggtggta gatggatcga
     4381 tggcaaacat gtcccagctc caagaaacga aaaggccgag atcgccatat ttggtgtccc
     4441 cgaggatcca aatttccaat cttctggtat taacttcgat aactacgatg atattccagt
     4501 ggacgcctct ggtaaggatg ttcctgaacc aatcacagaa tttacctcac ctccattgga
     4561 cggattgtta ttggaaaaca tcaaattggc ccgtttcacc aagccaacac ctgtgcaaaa
     4621 atactccgtc cctatcgttg ccaacggcag agatttgatg gcctgtgcgc agaccggttc
     4681 tggtaagact ggtgggtttt tattcccagt gttgtccgaa tcatttaaga ctggaccatc
     4741 tcctcaacca gagtctcaag gctcctttta ccaaagaaag gcctacccaa ctgctgtcat
     4801 tatggctcca gtttaaacca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4861 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4921 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4981 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5041 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5101 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5161 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5221 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5281 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5341 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5401 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5461 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5521 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5581 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5641 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5701 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5761 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5821 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5881 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5941 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6001 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6061 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6121 cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6181 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6241 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6301 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6361 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6421 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6481 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6541 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6601 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6661 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6721 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6781 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6841 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6901 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6961 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7021 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     7081 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7141 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7201 cagagcagat tgtactgaga gtgcaccata attcgtttaa accgttttaa gagcttggtg
     7261 agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg aagtcataac
     7321 acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag tacactctat
     7381 atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg gatgactctt
     7441 tttttttctt agcgattggc attatcacat aatgaattat acattatata aagtaatgtg
     7501 atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagatgac
//
