LOCUS       sequence-420132-        7295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A synthetic ABA signaling reporter designed by multimerization of 
            ABRE (7bp) and its flanking sequences from the ABI1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7295)
  AUTHORS   Wu R, Duan L, Pruneda-Paz JL, Oh DH, Pound M, Kay S, Dinneny JR
  TITLE     The 6xABRE Synthetic Promoter Enables the Spatiotemporal Analysis of
            ABA-Mediated Transcriptional Regulation.
  JOURNAL   Plant Physiol. 2018 Aug;177(4):1650-1665. doi: 10.1104/pp.18.00401. 
            Epub 2018 Jun 8.
  PUBMED    29884679
REFERENCE   2  (bases 1 to 7295)
  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.18.00401"; journalName: "Plant Physiol"; date: 
            "2018-08"; volume: "177"; issue: "4"; pages: "1650-1665"
COMMENT     Sequence Label: 6xABRE_A
FEATURES             Location/Qualifiers
     source          1..7295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     promoter        85..430
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1337..2047
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMEELYK"
     primer_bind     complement(1482..1500)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1750..1769
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2066..2318
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2563..2609
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2683..3393
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2710..2738)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3333..3353
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3470..3646
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(3710..3730)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3710..3728)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3749..3765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3749..3765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3762..3784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3773..3789
                     /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(3797..3803,3804..3821,3822..3827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3842..3863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3980..3997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4102..4126
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4217..4805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4297..4316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4896..5684)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6736..7171
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 cttggcagga tatattgtgg tgtaacgttg gcgcgcctac tccaaaaatg tcaaagatac
       61 agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaattt cgggaaacct
      121 cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag aaaaggaagg
      181 tggctcctac aaatgccatc attgcgataa aggaaaggct atcattcaag atgcctctgc
      241 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
      301 tccaaccacg tcttcaaagc aagtggattg atgtgacatc tccactgacg taagggatga
      361 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
      421 ggagaggaca gcccagatca ctagtcttaa gtccggagct agctctagaa tggcaaagga
      481 tgtggaagcc gttcccggag aaggatttca gacaagagac tatcaagatc cgccaccagc
      541 tccgtttatt gatggagcgg agctaaagaa gtggtctttc tacagagcag ttatcgcaga
      601 gttcgtagcc actctcctct tcttatacat caccgttttg acagtcatcg gttacaagat
      661 tcagtccgat actgatgccg gtggcgtaga ttgcggcgga gttggaatcc tcggtatcgc
      721 ttgggccttt ggtggtatga tcttcatcct cgtctactgc accgccggta tctctggtgg
      781 tcacattaac ccagcggtga catttgggct attcttggca cgtaaagtgt cgttacctag
      841 ggccctattg tacataatcg ctcagtgttt gggtgcgatt tgtggagttg gttttgtcaa
      901 agccttccaa agctcttact acacccgtta cggaggtgga gccaactctc tagccgatgg
      961 ctacagcaca gggaccggtc tagccgcaga gatcattggt actttcgttc ttgtctacac
     1021 cgtcttctct gccactgacc ccaaacgtag tgccagagac tcccacgttc cggtgttggc
     1081 gccacttcca atcggatttg ccgtgttcat ggtacatttg gctaccattc ccattaccgg
     1141 aaccggaatt aacccggcaa ggagtttcgg agctgccgta atctacaaca agagcaagcc
     1201 atgggatgac cactggatat tttgggttgg accattcatt ggagctgcga tagctgcatt
     1261 ctaccaccaa ttcgttctaa gagcttcagg ttctaagtct cttggatcat tcagaagtgc
     1321 tgccaacgtc ggatccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     1381 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     1441 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     1501 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     1561 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     1621 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1681 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1741 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     1801 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1861 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1921 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     1981 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg aggagctgta
     2041 caagtaaagc ggccgcccgg ctgcagatcg ttcaaacatt tggcaataaa gtttcttaag
     2101 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     2161 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
     2221 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
     2281 taaattatcg cgcgcggtgt catctatgtt actagatcgg ggcgcgccct cgagagtatt
     2341 ttcttcgtct acgtgtcgac catccacctt ttcttcgtct acgtgtcgac catccacctt
     2401 ttcttcgtct acgtgtcgac catccacctt ttcttcgtct acgtgtcgac catccacctt
     2461 ttcttcgtct acgtgtcgac catccacctt ttcttcgtct acgtgtcgac catccaccat
     2521 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     2581 ataaggaagt tcatttcatt tggagaggac acgctgatga agactaatct ttttctcttt
     2641 ctcatctttt cacttctcct atcattatcc tcggccgaat tcagtaaagg agaagaactt
     2701 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     2761 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     2821 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt ctcttatggt
     2881 gttcaatgct tttcaagata cccagatcat atgaagcggc acgacttctt caagagcgcc
     2941 atgcctgagg gatacgtgca ggagaggacc atcttcttca aggacgacgg gaactacaag
     3001 acacgtgctg aagtcaagtt tgagggagac accctcgtca acaggatcga gcttaaggga
     3061 atcgatttca aggaggacgg aaacatcctc ggccacaagt tggaatacaa ctacaactcc
     3121 cacaacgtat acatcatggc cgacaagcaa aagaacggca tcaaagccaa cttcaagacc
     3181 cgccacaaca tcgaagacgg cggcgtgcaa ctcgctgatc attatcaaca aaatactcca
     3241 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     3301 tcgaaagatc ccaacgaaaa gagagatcac atggtccttc ttgagtttgt aacagctgct
     3361 gggattacac atggcatgga tgaactatac aaacatgatg agctttaatg gaaagcttgt
     3421 cggtacgctg aaatcaccag tctctctcta caaatctatc tctctctatt ttctccataa
     3481 ataatgtgtg agtagtttcc cgataaggga aattagggtt cttatagggt ttcgctcatg
     3541 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     3601 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatcgatg ggctagagcg
     3661 gactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     3721 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3961 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4081 caggaaagaa catgaaggcc ttgacaggat atattggcgg gtaaactaag tcgctgtatg
     4141 tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4621 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4681 tgctgaagcc agttaccttc ggaagaagag ttggtagctc ttgatccggc aaacaaacca
     4741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4801 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4861 gttaagggat tttggtcatg aattcttgtt agacattatt tgccgactac cttggtgatc
     4921 tcgcctttca cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     4981 tcttgtccaa gataagcctg cctagcttca agtatgacgg gctgatactg ggccggcagg
     5041 cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg
     5101 taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc
     5161 gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga
     5221 tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc
     5281 agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca
     5341 ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg
     5401 tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa
     5461 gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg
     5521 gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag
     5581 ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc
     5641 tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgatgtt taactcctga
     5701 attaagccgc gccgcgaagc ggtgtcggct tgaatgaatt gttaggcgtc atcctgtgct
     5761 cccgagaacc agtaccagta catcgctgtt tcgttcgaga cttgaggtct agttttatac
     5821 gtgaacaggt caatgccgcc gagagtaaag ccacattttg cgtacaaatt gcaggcaggt
     5881 acattgttcg tttgtgtctc taatcgtatg ccaaggagct gtctgcttag tgcccacttt
     5941 ttcgcaaatt cgatgagact gtgcgcgact cctttgcctc ggtgcgtgtg cgacacaaca
     6001 atgtgttcga tagaggctag atcgttccat gttgagttga gttcaatctt cccgacaagc
     6061 tcttggtcga tgaatgtgcc atagcaagca gagtcttcat cagagtcatc atccgagatg
     6121 taatccttcc ggtaggggct cacacttctg gtagatagtt caaagccttg gtcggatagg
     6181 tgcacatcga acacttcacg aacaatgaaa tggttctcag catcaaatgt ttccgccacc
     6241 tgctcaggga tcaccgaaat cttcatatga cgcctaacgc ctggcacagc ggatcgcaaa
     6301 cctggcgcgg cttttggcac aaaaggcgtg acaggtttgc gaatccgttg ctgccacttg
     6361 tttaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6421 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat
     6481 tgcagcactg ccatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     6541 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     6601 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     6661 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     6721 cgattcagga gatccccatc caacagcccg ccgtcgagcg ggctttttta tccccggaag
     6781 cctgtggata gagggtagtt atccacgtga aaccgctaat gccccgcaaa gccttgattc
     6841 acggggcttt ccggcccgct ccaaaaacta tccacgtgaa atcgctaatc agggtacgtg
     6901 aaatcgctaa tcggagtacg tgaaatcgct aataaggtca cgtgaaatcg ctaatcaaaa
     6961 aggcacgtga gaacgctaat agccctttca gatcaacagc ttgcaaacac ccctcgctcc
     7021 ggcaagtagt tacagcaagt agtatgttca attagctttt caattatgaa tatatatatc
     7081 aattattggt cgcccttggc ttgtggacaa tgcgctacgc gcaccggctc cgcccgtgga
     7141 caaccgcaag cggttgccca ccgtcgagcg cctttgccca caacccggcg gccggccgca
     7201 acagatcgtt ttataaattt ttttttttga aaaagaaaaa gcccgaaagg cggcaacctc
     7261 tcgggcttct ggatttccga tccccggaat tagat
//
