LOCUS       sequence-59814-e        8654 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 8654)
  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 8654)
  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: pCambia1300-PacI
FEATURES             Location/Qualifiers
     source          1..8654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..28
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        219..895
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     832..852
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             962..1987
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    2027..2201
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2279..2303
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2728..3522
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3609..4197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4098..4117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4351..4368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4383..4523
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4449..4468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4609..4631
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4867..5061
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5127..6194)
                     /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"
     CDS             complement(6628..7257)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    8557..8581
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
       61 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      121 gcgtattggc tagagcagct tgccaacatg gtggagcacg acactctcgt ctactccaag
      181 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
      241 atgtcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
      301 gtagaaaggg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
      361 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
      421 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga acatggtgga
      481 gcacgacact ctcgtctact ccaagaatat caaagataca gtctcagagg accaaagggc
      541 tattgagact tttcaacaaa gggtaatgtc gggaaacctc ctcggattcc attgcccagc
      601 tatctgtcac ttcatcaaaa ggacagtaga aagggaaggt ggcacctaca aatgccatca
      661 ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg
      721 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
      781 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
      841 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctgaaatca
      901 ccagtctctc tctacaaatc tatctctctc gagctttcgc agatccgggg ggcaatgaga
      961 tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     1021 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     1081 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     1141 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     1201 tggggagttt agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     1261 gcaagacctg cctgaaaccg aactgcccgc tgttctacaa ccggtcgcgg aggccatgga
     1321 tgcaatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     1381 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     1441 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     1501 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     1561 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     1621 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     1681 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     1741 acgactccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     1801 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     1861 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     1921 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     1981 gaaatagagt agatgccgac cggatctgtc gatcgacaag ctcgagtttc tccataataa
     2041 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
     2101 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
     2161 tctaattcct aaaaccaaaa tccagtacta aaatccagat cccccgaatt aattcggcgt
     2221 taattcagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt
     2281 ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca
     2341 caaaatcacc actcgataca ggcagcccat cagtccggga cggcgtcagc gggagagccg
     2401 ttgtaaggcg gcagactttg ctcatgttac cgatgctatt cggaagaacg gcaactaagc
     2461 tgccgggttt gaaacacgga tgatctcgcg gagggtagca tgttgattgt aacgatgaca
     2521 gagcgttgct gcctgtgatc accgcggttt caaaatcggc tccgtcgata ctatgttata
     2581 cgccaacttt gaaaacaact ttgaaaaagc tgttttctgg tatttaaggt tttagaatgc
     2641 aaggaacagt gaattggagt tcgtcttgtt ataattagct tcttggggta tctttaaata
     2701 ctgtagaaaa gaggaaggaa ataataaatg gctaaaatga gaatatcacc ggaattgaaa
     2761 aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta
     2821 tataaactag tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa
     2881 gggaccacct atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg
     2941 cctgttccaa aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt
     3001 gaggccgatg gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt
     3061 atcgagctgt atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc
     3121 tatacgaata gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg
     3181 gccgatgtgg attgcgaaaa ctgggaagaa gacactccat ttaaagatcc gcgcgagctg
     3241 tatgattttt taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg
     3301 ggagacagca acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga
     3361 agcggcaggg cggacaagtg gtatgacatt gccttctgcg tccggtcgat cagggaggat
     3421 atcggggaag aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg
     3481 gagaaaataa aatattatat tttactggat gaattgtttt agtacctaga atgcatgacc
     3541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3721 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4321 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4381 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4441 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     4501 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4561 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4621 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg
     4681 cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt
     4741 aggccagcca tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt
     4801 agggagcgca gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca
     4861 gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag
     4921 gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca
     4981 atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc
     5041 ccaatgtacg tgctatccac aggaaagaga ccttttcgac ctttttcccc tgctagggca
     5101 atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg
     5161 ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac
     5221 tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct
     5281 ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct
     5341 gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa
     5401 tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa
     5461 tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag
     5521 cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc
     5581 gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct
     5641 ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg
     5701 aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga
     5761 tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct
     5821 gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg
     5881 tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc
     5941 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
     6001 atcgacggcg caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc
     6061 gcttgccacg attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc
     6121 gcggccttca acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg
     6181 gatggtttgc gaccgtcacg ccgattcctc gggcttgggg gttccagtgc cattgcaggg
     6241 ccggcagaca acccagccgc ttacgcctgg ccaaccgccc gttcctccac acatggggca
     6301 ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt tagccgctaa
     6361 aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg atgtattcag
     6421 atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc ttggtgtgat
     6481 cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc aggctggcca
     6541 acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt
     6601 tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt cagcggccag
     6661 cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc
     6721 ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc
     6781 gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga tcgcccgcga
     6841 cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct taaccagctc
     6901 caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa tcagcacgaa
     6961 gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc cgtcgatcac
     7021 tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc
     7081 gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg
     7141 atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg ctagatgggt
     7201 tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa ccttcatgcg
     7261 ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac cgcatgacgc
     7321 aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt ttcttcagcg
     7381 gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag gatttcatgc
     7441 agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc gatcatctcc
     7501 gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg
     7561 cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt
     7621 cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa
     7681 gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct ttcacggtgc
     7741 ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc
     7801 gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga
     7861 tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg
     7921 ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg ccggcctcct
     7981 ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg
     8041 tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc tccgggcggt
     8101 cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt
     8161 cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag cccagcaggc
     8221 cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa gtattctact
     8281 ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc
     8341 gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact gaacgtcaga
     8401 agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc cgaggggaac
     8461 cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa
     8521 atatccgtta ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc
     8581 aaacactgat agtttaaact gaaggcggga aacgacaatc tgatccaagc tcaagctgct
     8641 ctagcattaa ttaa
//
