LOCUS       sequence-167586-        5917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  target  miRNA394  for destruction in Arabidopsis thaliana.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5917)
  AUTHORS   Tang G, Yan J, Gu Y, Qiao M, Fan R, Mao Y, Tang X
  TITLE     Construction of short tandem target mimic (STTM) to block the 
            functions of plant and animal microRNAs.
  JOURNAL   Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006. 
            Epub 2012 Oct 23.
  PUBMED    23098881
REFERENCE   2  (bases 1 to 5917)
  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: 
            "10.1016/j.ymeth.2012.10.006"; journalName: "Methods"; date: 
            "2012-10"; volume: "58"; issue: "2"; pages: "118-25"
COMMENT     Sequence Label: pFGC5941-PacI-ath-STTM394
FEATURES             Location/Qualifiers
     source          1..5917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(365..384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(366..384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        577..957
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             963..1514
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      1524..1776
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    1862..1886
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2311..3105
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3192..3780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3681..3700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3842..3847,3848..3864,3865..3870)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3973..3992)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        4631..4649
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4631..4648
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    5211..5387
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(5530..5875)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 gagtaggggt gggattgtgc gtcatccctt acgtcagtgg agatgtcaca tcaacccact
       61 tgctttgaag acgtggttgg aacgtcttct tttcccacga tgctcctcgt gggtgggggt
      121 ccatctttgg gaccactgtc ggcagaggca tcttgaatga tagcctttcc tttatcgcaa
      181 tgatggcatt tgtaggagcc accttccttt tccactgtcc tttcgatgaa gtgacagata
      241 gctgggcaat ggaatccgag gaggtttccc gaaattaccc tttgttgaaa agtctcaata
      301 gccctttggt cttctgagac tgtatctttg acatttttgg agtagggatc agatctgccg
      361 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ctaatgaatc
      421 ggctgcagta cccgggaatt taacccgcct aatgagcggg cttttttttt tgtgatccaa
      481 aggatctcct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa cttaattaac
      541 tctctagact cacctaggat ccatgaattc tgagattttt caaatcagtg cgcaagacgt
      601 gacgtaagta tccgagtcag tttttatttt tctactaatt tggtcgttta tttcggcgtg
      661 taggacatgg caaccgggcc tgaatttcgc gggtattctg tttctattcc aactttttct
      721 tgatccgcag ccattaacga cttttgaata gatacgctga cacgccaagc ctcgctagtc
      781 aaaagtgtac caaacaacgc tttacagcaa gaacggaatg cgcgtgacgc tcgcggtgac
      841 gccatttcgc cttttcagaa atggataaat agccttgctt cctattatat cttcccaaat
      901 taccaataca ttacactagc atctgaattt cataaccaat ctcgatacac caaatcgaat
      961 cgatgagccc agaacgacgc ccggccgaca tccgccgtgc caccgaggcg gacatgccgg
     1021 cggtctgcac catcgtcaac cactacatcg agacaagcac ggtcaacttc cgtaccgagc
     1081 cgcaggaacc gcaggagtgg acggacgacc tcgtccgtct gcgggagcgc tatccctggc
     1141 tcgtcgccga ggtggacggc gaggtcgccg gcatcgccta cgcgggcccc tggaaggcac
     1201 gcaacgccta cgactggacg gccgagtcga ccgtgtacgt ctccccccgc caccagcgga
     1261 cgggactggg ctccacgctc tacacccacc tgctgaagtc cctggaggca cagggcttca
     1321 agagcgtggt cgctgtcatc gggctgccca acgacccgag cgtgcgcatg cacgaggcgc
     1381 tcggatatgc cccccgcggc atgctgcggg cggccggctt caagcacggg aactggcatg
     1441 acgtgggttt ctggcagctg gacttcagcc tgccggtacc gccccgtccg gtcctgcccg
     1501 tcaccgagat ttgactgcag aatcttggac tcccatgttg gcaaaggcaa ccaaacaaac
     1561 aatgaatgat ccgctcctgc atatggggcg gtttgagtat ttcaactgcc atttgggctg
     1621 aattgaagac atgctcctgt cagaaattcc gtgatcttac tcaatattca gtaatctcgg
     1681 ccaatatcct aaatgtgcgt ggctttatct gtctttgtat tgtttcatca attcatgtaa
     1741 cgtttgcttt tcttatgaat tttcaaataa attatcagat ccgagctcga attaattcgg
     1801 cgttaattca gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt
     1861 tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg
     1921 gcacaaaatc accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag
     1981 ccgttgtaag gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta
     2041 agctgccggg tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg
     2101 acagagcgtt gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt
     2161 atacgccaac tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa
     2221 tgcaaggaac agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa
     2281 atactgtaga aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg
     2341 aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag
     2401 gtatataagc tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat
     2461 aaagggacca cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag
     2521 ctgcctgttc caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg
     2581 agtgaggccg atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag
     2641 attatcgagc tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt
     2701 ccctatacga atagcttaga cagccgctta gccgaattgg attacttact gaataacgat
     2761 ctggccgatg tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag
     2821 ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac
     2881 ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg
     2941 agaagcggca gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag
     3001 gatatcgggg aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat
     3061 tgggagaaaa taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg
     3121 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3181 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3241 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3301 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3361 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3421 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3481 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3541 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3601 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3661 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3721 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3781 aacgccagca acgcggatca caacaaaaag cccgctcatt aggcgggcta aattctcatg
     3841 tttgacagct tatcatcgat aagctttaat gagttatcga gattttcagg agctaaggaa
     3901 gctaaaatgg agaaagaaat cactggatat accaccgttg atatatccca atggcatcgt
     3961 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     4021 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     4081 tttattcaca ttcttgcccg cctgatgaat gctcatccgg agttccgtat ggcaatgaaa
     4141 gacggtgagc tggtaatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     4201 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     4261 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     4321 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     4381 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     4441 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     4501 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg acagggcggg
     4561 gcgtaattgg tacgtcgacg taagaacgcg gccacaatta atacataacc ttatgtatca
     4621 tacacatacg atttaggtga cactatagaa ctcgagcagc tgaagctcca atgtgatggt
     4681 ctagaggtcg catgcgatcc ccgtcaccgg tgtgagggaa ctagttttga tcttgaaaga
     4741 tcttttatct ttagagttaa ggactctttc gtatttcggt gaggttttat cctcttgagt
     4801 tttggtcata gacctattca tggctctgat accaattttt aagcgggggc ttatgcggat
     4861 tatttcttaa attgataagg ggttattagg gggtataggg tataaataca atcattccct
     4921 tagcgtatag tataagtata gtagcgtacc tctatcaaat ttccatcttc ttaccttgca
     4981 cagggcctgc aaccttatcc ttccttgtct tcctccttcc ttccgtccac ttcatcatat
     5041 ttaaaccaaa cctacggggg agtcaacgta accaaccctg ccttagcatc ttttccctaa
     5101 cggcctcctg cctaagcggt acttctagct tcgaacggcg tctgggctcc aggtctagtc
     5161 gtctcgtgtc tggtttatat tcacgacaaa gatctatagg gactttagga gatctggatt
     5221 ttagtactgg attttggttt taggaattag aaattttatt gatagaagta ttttacaaat
     5281 acaaatacat actaagggtt tcttatatgc tcaacacatg agcgaaaccc tataagaacc
     5341 ctaatttccc ttatcgggaa actactcaca cattatttat ggagaaaata gagagagata
     5401 gatttgtaga gagagactgg tgatttcagc gtaccgaatt cttggcattc ttaggtccac
     5461 ctccattctt cttctttaga ccatatttac aacaacttgg cattcttagg tccacctcca
     5521 agcttgggct gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttacga
     5581 aggatagtgg ggttgtgcgt catcccttat gtcagtggag atgtcacatc aatccacttg
     5641 ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
     5701 atctttggga ccactgtcgg cagaggcatc ttgaatgata gcctttcctt tatcgcaatg
     5761 atggcatttg taggagccac cttccttttc tactgtcctt tcgatgaggt gacagatagc
     5821 tgggcaacgg aatccgagga ggtttcccga aattaccctt tgttgaaaag tctcaatagc
     5881 cctttggtct tctgagacag tatctttgac atttttg
//
