LOCUS       sequence-370250-        7565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabidopsis Nuclear Auxin Signaling Receptor AFB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7565)
  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 7565)
  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-AtAFB2
FEATURES             Location/Qualifiers
     source          1..7565
                     /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             3262..3327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3328..3345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3393..3517)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3550..3566)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3551..3567)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3569..3815
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3585..3603)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4829..4845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4829..4845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4842..4864)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4853..4869
                     /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(4877..4883,4884..4901,4902..4907))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4922..4943
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5060..5077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5231..5819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5311..5330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5990..6781,6782..6850))
                     /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     6613..6632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6851..6955)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7023..7041
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7079..7101)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7201..7220
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7373..7560
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(7561..7565,1..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 caaaatgaat tatttcccag atgaagtaat agagcatgta
     1561 ttcgactttg taacatctca caaagacagg aatgctatat ctcttgtatg caaatcatgg
     1621 tacaagattg aaagatacag taggcaaaag gttttcattg gaaactgtta tgccattaat
     1681 ccagagaggt tgcttcggag attcccatgt ctaaagtctt tgactttgaa aggaaaacct
     1741 cattttgcgg atttcaattt ggttcctcat gaatggggag gttttgtgct accttggatt
     1801 gaggctttgg ctagaagccg tgtaggactt gaagagctta ggttgaagag gatggttgtt
     1861 actgatgaga gtcttgagct gctttctcgt tcttttgtca attttaagtc tttggtcctt
     1921 gttagctgtg aaggttttac cactgatggt cttgcctcta ttgccgctaa ttgcaggcat
     1981 cttcgggatc ttgatttgca agagaatgaa atcgatgatc atagaggtca atggttaagt
     2041 tgtttcccag acacttgcac gactcttgtc acgctaaact ttgcgtgcct cgaaggagaa
     2101 actaatctgg ttgctctaga gaggcttgtt gctagatctc caaacctaaa gagtctgaag
     2161 ctaaatcgtg cagtaccgtt agatgcactc gcaaggttaa tggcgtgtgc gccgcagata
     2221 gttgacttag gagtagggtc ttatgagaat gacccagatt ccgagtctta cttgaaactc
     2281 atggctgtca taaagaaatg cacctcgttg aggagtttgt cgggttttct agaggctgct
     2341 cctcactgtc tctcagcttt ccacccaata tgtcataacc tcacctcctt gaatcttagt
     2401 tacgcagctg agattcatgg tagccacctt attaagctta ttcagcattg caagaaactt
     2461 cagcggttat ggattttgga tagtataggt gacaaagggc ttgaagttgt agcttctaca
     2521 tgtaaagagt tacaagagct tagggttttt ccatctgatt tactcggtgg aggcaacaca
     2581 gctgtgaccg aagaaggtct agttgccatc tcggcaggct gccctaagct ccactctata
     2641 ctctacttct gccaacaaat gacaaacgca gctctcgtaa ccgttgccaa gaactgtcca
     2701 aatttcatcc gtttccgact ctgcatcctc gagccaaaca aacccgatca cgtcacatct
     2761 caacctctag acgaaggctt tggagcaatc gtcaaagcct gcaagagcct gagaaggctt
     2821 tctctctcag gactccttac agaccaagtc ttcctctaca tcggaatgta cgcgaatcag
     2881 ctcgagatgc tctccatagc ctttgcagga gatacagaca aaggcatgct atatgtgttg
     2941 aatggttgca aaaagatgaa gaaactagag ataagggata gtccgtttgg ggacacggcg
     3001 cttcttgctg atgtgagcaa gtatgaaaca atgcgatccc tttggatgtc ttcatgtgaa
     3061 gtcacactca gtggatgcaa aaggctcgca gagaaagcgc catggctcaa tgtagagatc
     3121 ataaacgaga atgataataa ccggatggaa gaaaacggac acgaggggag gcagaaagtg
     3181 gataagttgt atctgtaccg gactgtggtt gggacaagaa tggatgcgcc gccatttgtg
     3241 tggattctcg gatctggtgg agattacaag gatcatgatg gtgattacaa ggatcacgac
     3301 atcgactaca aggatgacga tgacaagcac catcatcacc accattgatt ctatgtcagg
     3361 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     3421 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3481 tttacgtttc tcgttcagct ttcttgtaca aagtggtgat gggctgcagg aattcgattt
     3541 caagcttatc gataccgtcg acctcgagtc atgtaattag ttatgtcacg cttacattca
     3601 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     3661 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     3721 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     3781 gagaaggttt tgggacgctc gaaggcttta atttgtgaca ccgattattt aaagctgcag
     3841 catacgatat atatacatgt gtatatatgt atacctatga atgtcagtaa gtatgtatac
     3901 gaacagtatg atactgaaga tgacaaggta atgcatcatt ctatacgtgt cattctgaac
     3961 gaggcgcgct ttcctttttt ctttttgctt tttctttttt tttctcttga actcgagaaa
     4021 aaaaatataa aagagatgga ggaacgggaa aaagttagtt gtggtgatag gtggcaagtg
     4081 gtattccgta agaacaacaa gaaaagcatt tcatattatg gctgaactga gcgaacaagt
     4141 gcaaaattta agcatcaacg acaacaacga gaatggttat gttcctcctc acttaagagg
     4201 aaaaccaaga agtgccagaa ataacagtag caactacaat aacaacaacg gcggctacaa
     4261 cggtggccgt ggcggtggca gcttctttag caacaaccgt cgtggtggtt acggcaacgg
     4321 tggtttcttc ggtggaaaca acggtggcag cagatctaac ggccgttctg gtggtagatg
     4381 gatcgatggc aaacatgtcc cagctccaag aaacgaaaag gccgagatcg ccatatttgg
     4441 tgtccccgag gatccaaatt tccaatcttc tggtattaac ttcgataact acgatgatat
     4501 tccagtggac gcctctggta aggatgttcc tgaaccaatc acagaattta cctcacctcc
     4561 attggacgga ttgttattgg aaaacatcaa attggcccgt ttcaccaagc caacacctgt
     4621 gcaaaaatac tccgtcccta tcgttgccaa cggcagagat ttgatggcct gtgcgcagac
     4681 cggttctggt aagactggtg ggtttttatt cccagtgttg tccgaatcat ttaagactgg
     4741 accatctcct caaccagagt ctcaaggctc cttttaccaa agaaaggcct acccaactgc
     4801 tgtcattatg gctccagttt aaaccatggt catagctgtt tcctgtgtga aattgttatc
     4861 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4921 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4981 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5041 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5101 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5161 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5221 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5281 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5341 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5401 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5461 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5521 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5581 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5641 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5701 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5761 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5821 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5881 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5941 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6001 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6061 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6121 tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6181 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6241 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6301 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6361 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6421 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6481 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6541 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6601 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6661 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6721 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6781 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6841 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6901 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6961 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7021 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     7081 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     7141 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     7201 cggcatcaga gcagattgta ctgagagtgc accataattc gtttaaaccg ttttaagagc
     7261 ttggtgagcg ctaggagtca ctgccaggta tcgtttgaac acggcattag tcagggaagt
     7321 cataacacag tcctttcccg caattttctt tttctattac tcttggcctc ctctagtaca
     7381 ctctatattt ttttatgcct cggtaatgat tttcattttt ttttttccac ctagcggatg
     7441 actctttttt tttcttagcg attggcatta tcacataatg aattatacat tatataaagt
     7501 aatgtgattt cttcgaagaa tatactaaaa aatgagcagg caagataaac gaaggcaaag
     7561 atgac
//
