LOCUS       sequence-104553-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The artificial microRNA is designed for down-regulating expression 
            of ACD6 mRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Todesco M, Balasubramanian S, Hu TT, Traw MB, Horton M, Epple P, 
            Kuhns C, Sureshkumar S, Schwartz C, Lanz C, Laitinen RA, Huang Y, 
            Chory J, Lipka V, Borevitz JO, Dangl JL, Bergelson J, Nordborg M, 
            Weigel D
  TITLE     Natural allelic variation underlying a major fitness trade-off in 
            Arabidopsis thaliana.
  JOURNAL   Nature. 2010 Jun 3;465(7298):632-6. doi: 10.1038/nature09083.
  PUBMED    20520716
REFERENCE   2  (bases 1 to 6580)
  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"; journalName: 
            "Nature"; date: "2010-06-3- 3"; volume: "465"; issue: "7298"; pages:
            "632-6"
COMMENT     Sequence Label: MT79
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    542..564
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     terminator      complement(588..839)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(859..1410)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIQTSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1453..1632)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1864..1886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1878..1895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1879..1895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1905..1924
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1905..1923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2454..2799
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2742..2762
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2821..2845
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2859..2875
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2860..2876
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3316..3332)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3358..3382)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3390..3406)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3410..4117
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(4150..4168)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4151..4168)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4207..4227)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4207..4225)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4246..4262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4246..4262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4259..4281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4270..4286
                     /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(4294..4300,4301..4318,4319..4324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4339..4360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4477..4494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4599..4623
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4714..5302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4794..4813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5473..6261)
                     /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      join(6552..6580,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgccag cgcctttgcc
      421 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
      481 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagagatc
      541 ttggcaggat atattgtggt gtaacgttat cagcttgcat gccggtcgat ctagtaacat
      601 agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc
      661 gtattaaatg tataattgcg ggactctaat caaaaaaccc atctcataaa taacgtcatg
      721 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
      781 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
      841 gcttgactct aggggtcatc agatttcggt gacgggcagg accggacggg gcggcaccgg
      901 caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct tgaagccggc
      961 cgcccgcagc atgccacggg gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc
     1021 gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca gggacttcag
     1081 caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg agacgtacac
     1141 ggttgactcg gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat
     1201 gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc gcagacggac
     1261 gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga ccgtgcttgt
     1321 ctggatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg tggcacggcg
     1381 gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc cctcgatcga gttgagagtg
     1441 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg
     1501 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     1561 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt ggctccttca
     1621 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     1681 cataacgtga ctcccttaat tctccgctca tgatcgataa cattaacgct tacaatttcc
     1741 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1801 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1861 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     1921 agggcgaatt gggtaccggg ccccccctcg atatgattcg gtccccagat tagccttttc
     1981 aatttcagaa agaatgctaa cccacagatg gttagagagg cttacgcagc aggtctcatc
     2041 aagacgatct acccgagcaa taatctccag gaaatcaaat accttcccaa gaaggttaaa
     2101 gatgcagtca aaagattcag gactaactgc atcaagaaca cagagaaaga tatatttctc
     2161 aagatcagaa gtactattcc agtatggacg attcaaggct tgcttcacaa accaaggcaa
     2221 gtaatagaga ttggagtctc taaaaaggta gttcccactg aatcaaaggc catggagtca
     2281 aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca gttcatacag
     2341 agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga gcacgacaca
     2401 cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc aattgagact
     2461 tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc tatctgtcac
     2521 tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca ttgcgataaa
     2581 ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc
     2641 acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
     2701 tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct
     2761 tcctctatat aaggaagttc atttcatttg gagagaacac gggggacgag ctcgcccatc
     2821 acaagtttgt acaaaaaagc aggctttcgg gccccccctc gaggtcgacg gtatcgataa
     2881 gcttgatatc gaattcctgc agccccaaac acacgctcgg acgcatatta cacatgttca
     2941 tacacttaat actcgctgtt ttgaattgat gttttaggaa tatatatgta gaacagcctt
     3001 tagtctccat tattcacagg tcgtgatatg attcaattag cttccgactc attcatccaa
     3061 ataccgagtc gccaaaattc aaactagact cgttaaatga atgaatgatg cggtagacaa
     3121 attggatcat tgattctctt tgattaatgg tgactaaagg ccgttctctc ttttgtattc
     3181 caattttctt gattaatctt tcctgcacaa aaacatgctt gatccactaa gtgacatata
     3241 tgctgccttc gtatatatag ttctggtaaa attaacattt tgggtttatc tttatttaag
     3301 gcatcgccat ggggggatcc actagttcta gagcggccgc caccgcggtg gatctagacc
     3361 cagctttctt gtacaaagtg gtgatggggg atccactagt tctagagtcc tgctttaatg
     3421 agatatgcga gacgcctatg atcgcatgat atttgctttc aattctgttg tgcacgttgt
     3481 aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca ttctaatgaa
     3541 tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat ttactgattg
     3601 taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc tgaataggtt
     3661 tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta ttattacaaa
     3721 tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta cataaatctt
     3781 attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc ggcgaactaa
     3841 taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag attccttgaa
     3901 gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc cggtccagca
     3961 cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt tggtgtatgt
     4021 gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc
     4081 agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac cggcatgcaa gctagcttac
     4141 tagtgatatt ctatagtgtc acctaaatct gcggccgcca ccgcggtgga gctccagctt
     4201 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4261 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4321 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4441 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4561 agaatcaggg gataacgcag gaaagaacat gaaggccttg acaggatata ttggcgggta
     4621 aactaagtcg ctgtatgtgt ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc
     4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga agaagagttg gtagctcttg
     5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgtgtaaca
     5461 ttggtctagt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtgaacaaa
     5521 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tgtccaagat aagcctgcct
     5581 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     5641 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     5701 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     5761 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     5821 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     5881 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     5941 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     6001 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     6061 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     6121 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     6181 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     6241 ggcgatcacc gcttccctca taacacccct tgtattactg tttatgtaag cagacagttt
     6301 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     6361 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     6421 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     6481 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     6541 tcaggcgatc cccatccaac agcccgccgt cgagcgggct
//
