LOCUS       sequence-308247-        8610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8610)
  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 8610)
  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
FEATURES             Location/Qualifiers
     source          1..8610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(111..363)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(373..924)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(930..1310)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     terminator      1380..2087
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(2112..2129)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2112..2128)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2121..2143)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    2331..2355
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3656..4285
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4714..5787
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5853..6047
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6283..6305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6391..6531
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6446..6465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6546..6563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6717..7305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6797..6816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7392..8186)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcga gctcggatct gataatttat
      121 ttgaaaattc ataagaaaag caaacgttac atgaattgat gaaacaatac aaagacagat
      181 aaagccacgc acatttagga tattggccga gattactgaa tattgagtaa gatcacggaa
      241 tttctgacag gagcatgtct tcaattcagc ccaaatggca gttgaaatac tcaaaccgcc
      301 ccatatgcag gagcggatca ttcattgttt gtttggttgc ctttgccaac atgggagtcc
      361 aagattctgc agtcaaatct cggtgacggg caggaccgga cggggcggta ccggcaggct
      421 gaagtccagc tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg
      481 cagcatgccg cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg
      541 cagcccgatg acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg
      601 ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg ggggagacgt acacggtcga
      661 ctcggccgtc cagtcgtagg cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc
      721 gacctcgccg tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc
      781 gtccgtccac tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat
      841 gtagtggttg acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc
      901 ggccgggcgt cgttctgggc tcatcgattc gatttggtgt atcgagattg gttatgaaat
      961 tcagatgcta gtgtaatgta ttggtaattt gggaagatat aataggaagc aaggctattt
     1021 atccatttct gaaaaggcga aatggcgtca ccgcgagcgt cacgcgcatt ccgttcttgc
     1081 tgtaaagcgt tgtttggtac acttttgact agcgaggctt ggcgtgtcag cgtatctatt
     1141 caaaagtcgt taatggctgc ggatcaagaa aaagttggaa tagaaacaga atacccgcga
     1201 aattcaggcc cggttgccat gtcctacacg ccgaaataaa cgaccaaatt agtagaaaaa
     1261 taaaaactga ctcggatact tacgtcacgt cttgcgcact gatttgaaaa atctcagaat
     1321 tcatggatcc taggtgagtc tagagagtta attaagaccc gggactagtc cctagagtcc
     1381 tgctttaatg agatatgcga gacgcctatg atcgcatgat atttgctttc aattctgttg
     1441 tgcacgttgt aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca
     1501 ttctaatgaa tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat
     1561 ttactgattg taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc
     1621 tgaataggtt tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta
     1681 ttattacaaa tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta
     1741 cataaatctt attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc
     1801 ggcgaactaa taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag
     1861 attccttgaa gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc
     1921 cggtccagca cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt
     1981 tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt
     2041 gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac ctgcaggcat
     2101 gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2161 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2221 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcta gagcagcttg
     2281 agcttggatc agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt tgacaggata
     2341 tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg
     2401 gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca gggttcccct
     2461 cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg cttctgacgt
     2521 tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct
     2581 gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat
     2641 acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg
     2701 ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac
     2761 gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg
     2821 gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta
     2881 gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc
     2941 ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc
     3001 atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc
     3061 acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc
     3121 ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg
     3181 aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc
     3241 agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg
     3301 accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac
     3361 cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga
     3421 tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg ctgcatgaaa
     3481 tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag
     3541 aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg
     3601 cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa
     3661 ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct
     3721 agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg
     3781 cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga
     3841 ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga
     3901 cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct
     3961 gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt
     4021 taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc
     4081 gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc
     4141 cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac
     4201 aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc
     4261 tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca
     4321 aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc
     4381 ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc
     4441 aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac
     4501 atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc
     4561 taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat
     4621 gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa
     4681 tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg
     4741 gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc
     4801 gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct
     4861 gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag
     4921 ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc
     4981 cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga
     5041 gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg
     5101 gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc
     5161 gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca
     5221 cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac
     5281 ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag
     5341 gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag
     5401 atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg
     5461 taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt
     5521 ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag
     5581 gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc
     5641 aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat
     5701 ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc
     5761 gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta
     5821 gcaggggaaa aaggtcgaaa aggtctcttt cctgtggata gcacgtacat tgggaaccca
     5881 aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac
     5941 cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac
     6001 tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag
     6061 cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc
     6121 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
     6181 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
     6241 caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6301 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6361 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     6421 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     6481 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7141 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7381 cattctaggt actaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca
     7441 ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc
     7501 ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca
     7561 agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg
     7621 ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg
     7681 cgcggatctt taaatggagt gtcttcttcc cagttttcgc aatccacatc ggccagatcg
     7741 ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa
     7801 tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt
     7861 tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg
     7921 agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt
     7981 ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac
     8041 cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta
     8101 gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat
     8161 tccggtgata ttctcatttt agccatttat tatttccttc ctcttttcta cagtatttaa
     8221 agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc cttgcattct
     8281 aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg gcgtataaca
     8341 tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg ctctgtcatc
     8401 gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt
     8461 tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct
     8521 cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     8581 ctgccggtcg gggagctgtt ggctggctgg
//
