LOCUS       sequence-420134-        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 RD29A 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_R
FEATURES             Location/Qualifiers
     source          1..7295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    680..704
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     809..826
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    943..964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(979..984,985..1002,1003..1009)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1017..1033
                     /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     1022..1044
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1041..1057
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1041..1057
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1076..1096
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1078..1096
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1160..1336
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1413..2123)
                     /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(1453..1473)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2068..2096
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(2197..2243)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      complement(2488..2740)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2759..3469)
                     /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(3037..3056)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3306..3324
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(4376..4721)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4781..4803
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      4930..5365
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6417..7205
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt cttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga gatctcaaac
      661 aaacacatac agcgacttag tttacccgcc aatatatcct gtcaaggcct tcatgttctt
      721 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      781 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      841 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      901 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      961 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     1021 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     1081 taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc gctctagaac
     1141 tagtccgctc tagcccatcg atctggattt tagtactgga ttttggtttt aggaattaga
     1201 aattttattg atagaagtat tttacaaata caaatacata ctaagggttt cttatatgct
     1261 caacacatga gcgaaaccct ataagaaccc taatttccct tatcgggaaa ctactcacac
     1321 attatttatg gagaaaatag agagagatag atttgtagag agagactggt gatttcagcg
     1381 taccgacaag ctttccatta aagctcatca tgtttgtata gttcatccat gccatgtgta
     1441 atcccagcag ctgttacaaa ctcaagaagg accatgtgat ctctcttttc gttgggatct
     1501 ttcgaaaggg cagattgtgt ggacaggtaa tggttgtctg gtaaaaggac agggccatcg
     1561 ccaattggag tattttgttg ataatgatca gcgagttgca cgccgccgtc ttcgatgttg
     1621 tggcgggtct tgaagttggc tttgatgccg ttcttttgct tgtcggccat gatgtatacg
     1681 ttgtgggagt tgtagttgta ttccaacttg tggccgagga tgtttccgtc ctccttgaaa
     1741 tcgattccct taagctcgat cctgttgacg agggtgtctc cctcaaactt gacttcagca
     1801 cgtgtcttgt agttcccgtc gtccttgaag aagatggtcc tctcctgcac gtatccctca
     1861 ggcatggcgc tcttgaagaa gtcgtgccgc ttcatatgat ctgggtatct tgaaaagcat
     1921 tgaacaccat aagagaaagt agtgacaagt gttggccatg gaacaggtag ttttccagta
     1981 gtgcaaataa atttaagggt aagttttccg tatgttgcat caccttcacc ctctccactg
     2041 acagaaaatt tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca
     2101 gtgaaaagtt cttctccttt actgaattcg gccgaggata atgataggag aagtgaaaag
     2161 atgagaaaga gaaaaagatt agtcttcatc agcgtgtcct ctccaaatga aatgaacttc
     2221 cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag
     2281 tggagataga gataaaggga cacgtatgaa gcgtctgaga gataaaggga cacgtatgaa
     2341 gcgtctgaga gataaaggga cacgtatgaa gcgtctgaga gataaaggga cacgtatgaa
     2401 gcgtctgaga gataaaggga cacgtatgaa gcgtctgaga gataaaggga cacgtatgaa
     2461 gcgtctgtac tctcgagggc gcgccccgat ctagtaacat agatgacacc gcgcgcgata
     2521 atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg
     2581 ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta
     2641 catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat
     2701 tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcagccggg cggccgcttt
     2761 acttgtacag ctcctccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt
     2821 ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg
     2881 cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt
     2941 ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg
     3001 ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt
     3061 cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg
     3121 aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact
     3181 tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt
     3241 aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc
     3301 cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct
     3361 cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca
     3421 tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg gatccgacgt
     3481 tggcagcact tctgaatgat ccaagagact tagaacctga agctcttaga acgaattggt
     3541 ggtagaatgc agctatcgca gctccaatga atggtccaac ccaaaatatc cagtggtcat
     3601 cccatggctt gctcttgttg tagattacgg cagctccgaa actccttgcc gggttaattc
     3661 cggttccggt aatgggaatg gtagccaaat gtaccatgaa cacggcaaat ccgattggaa
     3721 gtggcgccaa caccggaacg tgggagtctc tggcactacg tttggggtca gtggcagaga
     3781 agacggtgta gacaagaacg aaagtaccaa tgatctctgc ggctagaccg gtccctgtgc
     3841 tgtagccatc ggctagagag ttggctccac ctccgtaacg ggtgtagtaa gagctttgga
     3901 aggctttgac aaaaccaact ccacaaatcg cacccaaaca ctgagcgatt atgtacaata
     3961 gggccctagg taacgacact ttacgtgcca agaatagccc aaatgtcacc gctgggttaa
     4021 tgtgaccacc agagataccg gcggtgcagt agacgaggat gaagatcata ccaccaaagg
     4081 cccaagcgat accgaggatt ccaactccgc cgcaatctac gccaccggca tcagtatcgg
     4141 actgaatctt gtaaccgatg actgtcaaaa cggtgatgta taagaagagg agagtggcta
     4201 cgaactctgc gataactgct ctgtagaaag accacttctt tagctccgct ccatcaataa
     4261 acggagctgg tggcggatct tgatagtctc ttgtctgaaa tccttctccg ggaacggctt
     4321 ccacatcctt tgccattcta gagctagctc cggacttaag actagtgatc tgggctgtcc
     4381 tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt
     4441 gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt gaagacgtgg
     4501 ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac
     4561 tgtcggcaga ggcatcttga atgatagcct ttcctttatc gcaatgatgg catttgtagg
     4621 agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc
     4681 cgaggaggtt tcccgaaatt accctttgtt gaaaagtctc aatagccctt tggtcttctg
     4741 agactgtatc tttgacattt ttggagtagg cgcgccaacg ttacaccaca atatatcctg
     4801 ccaagatcta attccgggga tcggaaatcc agaagcccga gaggttgccg cctttcgggc
     4861 tttttctttt tcaaaaaaaa aaatttataa aacgatctgt tgcggccggc cgccgggttg
     4921 tgggcaaagg cgctcgacgg tgggcaaccg cttgcggttg tccacgggcg gagccggtgc
     4981 gcgtagcgca ttgtccacaa gccaagggcg accaataatt gatatatata ttcataattg
     5041 aaaagctaat tgaacatact acttgctgta actacttgcc ggagcgaggg gtgtttgcaa
     5101 gctgttgatc tgaaagggct attagcgttc tcacgtgcct ttttgattag cgatttcacg
     5161 tgaccttatt agcgatttca cgtactccga ttagcgattt cacgtaccct gattagcgat
     5221 ttcacgtgga tagtttttgg agcgggccgg aaagccccgt gaatcaaggc tttgcggggc
     5281 attagcggtt tcacgtggat aactaccctc tatccacagg cttccgggga taaaaaagcc
     5341 cgctcgacgg cgggctgttg gatggggatc tcctgaatcg ccccatcatc cagccagaaa
     5401 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
     5461 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     5521 tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac
     5581 attgcacaag ataaaaatat atcatcatgg cagtgctgca atgataccgc gagatccacg
     5641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5701 tggtcctgca actttatccg cctccatcca gtctattaaa caagtggcag caacggattc
     5761 gcaaacctgt cacgcctttt gtgccaaaag ccgcgccagg tttgcgatcc gctgtgccag
     5821 gcgttaggcg tcatatgaag atttcggtga tccctgagca ggtggcggaa acatttgatg
     5881 ctgagaacca tttcattgtt cgtgaagtgt tcgatgtgca cctatccgac caaggctttg
     5941 aactatctac cagaagtgtg agcccctacc ggaaggatta catctcggat gatgactctg
     6001 atgaagactc tgcttgctat ggcacattca tcgaccaaga gcttgtcggg aagattgaac
     6061 tcaactcaac atggaacgat ctagcctcta tcgaacacat tgttgtgtcg cacacgcacc
     6121 gaggcaaagg agtcgcgcac agtctcatcg aatttgcgaa aaagtgggca ctaagcagac
     6181 agctccttgg catacgatta gagacacaaa cgaacaatgt acctgcctgc aatttgtacg
     6241 caaaatgtgg ctttactctc ggcggcattg acctgttcac gtataaaact agacctcaag
     6301 tctcgaacga aacagcgatg tactggtact ggttctcggg agcacaggat gacgcctaac
     6361 aattcattca agccgacacc gcttcgcggc gcggcttaat tcaggagtta aacatcatga
     6421 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     6481 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     6541 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     6601 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     6661 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     6721 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     6781 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     6841 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     6901 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     6961 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     7021 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     7081 ccgtcatact tgaagctagg caggcttatc ttggacaaga agatcgcttg gcctcgcgcg
     7141 cagatcagtt ggaagaattt gttcactacg tgaaaggcga gatcaccaag gtagtcggca
     7201 aataatgtct aacaagaatt catgaccaaa atcccttaac gtgagttttc gttccactga
     7261 gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
