LOCUS       sequence-59592-e        8608 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 8608)
  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 8608)
  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..8608
                     /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             3655..4284
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4718..5785
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      5851..6045
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6281..6303)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6389..6529
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6444..6463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6544..6561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6715..7303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6795..6814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7390..8184)
                     /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 agttagatct 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 taaaaactag ctcagatact 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 ataacggata tttaaaaggg
     2401 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc
     2461 gggatcaaag tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt
     2521 cagtgcagcc gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg
     2581 ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata
     2641 cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg
     2701 gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg
     2761 cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg
     2821 agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag
     2881 accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg
     2941 gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca
     3001 tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca
     3061 cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc
     3121 tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga
     3181 aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca
     3241 gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga
     3301 ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc
     3361 gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat
     3421 cgcggccggg tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat
     3481 cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga
     3541 aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc
     3601 ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag
     3661 gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta
     3721 gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc
     3781 agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac
     3841 cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac
     3901 ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg
     3961 attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt
     4021 aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg
     4081 atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc
     4141 attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca
     4201 accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct
     4261 gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa
     4321 acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc
     4381 tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca
     4441 agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca
     4501 tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct
     4561 aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg
     4621 tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg gccctgcaat
     4681 ggcactggaa cccccaagcc cgaggaatcg gcgtgacggt cgcaaaccat ccggcccggt
     4741 acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc
     4801 ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga
     4861 tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc
     4921 gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg
     4981 cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc
     5041 tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc
     5101 cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga
     5161 atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca
     5221 cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct
     5281 ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc
     5341 caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat
     5401 cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta
     5461 ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt
     5521 gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc
     5581 agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa
     5641 gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt
     5701 gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga
     5761 gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc
     5821 aggggaaaaa ggtcgaaaag gtctctttcc tgtggatagc acgtacattg ggaacccaaa
     5881 gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg
     5941 gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc
     6001 tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg
     6061 cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc
     6121 cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg
     6181 caatctacca gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca
     6241 aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6301 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6361 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     6421 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6481 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     6541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6901 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7141 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7321 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca
     7381 ttctaggtac taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     7441 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     7501 gatcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     7561 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     7621 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     7681 cggatcttta aatggagtgt cttcttccca gttttcgcaa tccacatcgg ccagatcgtt
     7741 attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc
     7801 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc
     7861 agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag
     7921 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     7981 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     8041 gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat ataccttagc
     8101 aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc
     8161 cggtgatatt ctcattttag ccatttatta tttccttcct cttttctaca gtatttaaag
     8221 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     8281 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata
     8341 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt
     8401 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
     8461 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     8521 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     8581 gccggtcggg gagctgttgg ctggctgg
//
