LOCUS       sequence-240988-        6619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector containing 35S:MS2-p300:tRBCS followed by 
            GreenGate A-G sequences for insertion of additional modules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  AUTHORS   Lee JE, Neumann M, Duro DI, Schmid M
  TITLE     CRISPR-based tools for targeted transcriptional and epigenetic 
            regulation in plants.
  JOURNAL   PLoS One. 2019 Sep 26;14(9):e0222778. doi: 
            10.1371/journal.pone.0222778. eCollection 2019.
  PUBMED    31557222
REFERENCE   2  (bases 1 to 6619)
  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"; journalName: "PLoS One.";
            date: "2019-09-26"; volume: "14(9):e0222778. doi"; pages: " 
            10.1371/journal.pone.0222778. eCollection 2019"
COMMENT     Sequence Label: pJL024
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..541
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     484..504
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             592..978
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1033..1053
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      2938..3579
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    3685..3709
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3800..4388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3880..3899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4559..5347)
                     /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      5638..6073
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    6200..6222
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 tgaatcaaag gccatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa
       61 gactggcgaa cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt
      121 caacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga
      181 agaccaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt
      241 ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta
      301 caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg
      361 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      421 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      481 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac
      541 acgggggacg agctaacagt attcagtcga ctggtaccaa caggctccat ggcttctaat
      601 tttactcaat ttgttttggt tgataatgga ggaactggag atgttactgt tgctccttct
      661 aattttgcta atggagttgc tgaatggatt tcttctaatt caagatctca ggcttataag
      721 gttacttgtt ctgttagaca atcttctgct caaaagagaa agtatactat taaggttgaa
      781 gttcctaagg ttgctactca aactgttgga ggagttgaat tgcctgttgc tgcttggaga
      841 tcttatttga atatggaatt gactattcct atttttgcta ctaattctga ttgtgaattg
      901 attgttaagg ctatgcaagg attgttgaag gatggaaatc ctattccttc tgctattgct
      961 gctaattctg gaatttattc tgctggagga ggaggatctg gaggaggagg atctggagga
     1021 ggaggatctg gacctaagaa gaagagaaag gttgctgctg ctggatcttc aggtattttt
     1081 aagccagagg agcttagaca agcacttatg cctactcttg aggctcttta cagacaagat
     1141 cctgaatctc ttccattcag acagcctgtg gaccctcagc ttcttggtat tcctgactac
     1201 ttcgatattg tgaagtctcc tatggatctt tctacaatta aaagaaaact tgacacagga
     1261 caataccaag aaccttggca atacgttgat gacatctggc ttatgtttaa caatgcttgg
     1321 ctttacaaca gaaagacttc tagagtttac aaatattgct ctaaactttc tgaagtgttc
     1381 gaacaagaaa ttgatccagt gatgcaatct cttggttact gttgcggtag aaaacttgag
     1441 ttctctcctc aaactctttg ctgctatgga aagcaacttt gtactattcc tagagatgca
     1501 acatactatt cttatcaaaa cagataccat ttctgtgaaa aatgctttaa cgaaatccag
     1561 ggtgaatctg tttctcttgg agacgaccct tctcagcctc agacaacaat caataaagag
     1621 cagttctcta agagaaagaa cgatactctt gaccctgagc ttttcgtgga gtgtactgag
     1681 tgcggtagaa agatgcatca gatttgtgtt cttcatcacg agatcatttg gccagctggt
     1741 tttgtttgcg atggttgtct taagaagtct gcaagaacta gaaaagagaa taagttctct
     1801 gcaaagagac ttccttctac tagacttgga actttccttg aaaacagagt gaacgatttc
     1861 cttagaagac agaatcatcc tgagtctgga gaggtgacag tgagagtggt tcatgcatct
     1921 gataaaactg ttgaggttaa acctggaatg aaagctagat tcgtggactc tggagagatg
     1981 gcagaatctt ttccatacag aacaaaagct ctttttgcat tcgaagagat tgatggtgtt
     2041 gatctttgtt tcttcggtat gcacgtgcaa gagtatggat ctgactgccc tccaccaaat
     2101 caaagaagag tttacatttc ttatcttgat tctgttcact tttttagacc aaaatgcctt
     2161 agaacagcag tgtaccacga gattcttatt ggttatcttg agtatgtgaa aaaacttggt
     2221 tacacaactg gacatatttg ggcttgccca ccttctgaag gtgatgatta tatttttcat
     2281 tgccaccctc cagaccaaaa gattcctaaa cctaaaagac ttcaagagtg gtacaaaaag
     2341 atgcttgata aagcagtgtc tgagagaatt gtgcacgact acaaggacat ttttaaacag
     2401 gctacagagg atagacttac ttctgcaaaa gagcttcctt attttgaagg agatttttgg
     2461 cctaatgttc ttgaggagtc tattaaggaa cttgagcagg aggaagaaga gagaaaaaga
     2521 gaagagaaca catctaatga gtctacagac gttactaaag gtgattctaa aaacgctaaa
     2581 aagaaaaaca ataaaaaaac ttctaaaaat aaatcttctc tttctagagg taacaaaaaa
     2641 aaacctggaa tgccaaacgt ttctaatgac ctttctcaaa aactttacgc aacaatggaa
     2701 aaacataaag aagttttctt cgtgatcaga cttattgctg gacctgctgc taattctctt
     2761 cctccaatcg ttgacccaga ccctcttatt ccttgtgatc ttatggatgg aagagacgca
     2821 tttcttactc ttgcaagaga caaacatctt gaattttctt ctcttagaag agcacagtgg
     2881 tctacaatgt gtatgcttgt tgagcttcac acacaatctc aagactagta actgcaaagc
     2941 tttcgttcgt atcatcggtt tcgacaacgt tcgtcaagtt caatgcatca gtttcattgc
     3001 gcacacacca gaatcctact gagtttgagt attatggcat tgggaaaact gtttttcttg
     3061 taccatttgt tgtgcttgta atttactgtg ttttttattc ggttttcgct atcgaactgt
     3121 gaaatggaaa tggatggaga agagttaatg aatgatatgg tccttttgtt cattctcaaa
     3181 ttaatattat ttgttttttc tcttatttgt tgtgtgttga atttgaaatt ataagagata
     3241 tgcaaacatt ttgttttgag taaaaatgtg tcaaatcgtg gcctctaatg accgaagtta
     3301 atatgaggag taaaacactt gtagttgtac cattatgctt attcactagg caacaaatat
     3361 attttcagac ctagaaaagc tgcaaatgtt actgaataca agtatgtcct cttgtgtttt
     3421 agacatttat gaactttcct ttatgtaatt ttccagaatc cttgtcagat tctaatcatt
     3481 gctttataat tatagttata ctcatggatt tgtagttgag tatgaaaata ttttttaatg
     3541 cattttatga cttgccaatt gattgacaac atgcatcaat cgactatgag taggcaagat
     3601 gttctgtagg accttgagac cgaaaaggtg gtctcagtat tctagaccaa gcatgcacag
     3661 ctggcgttct cagccagatc cccttgacag gatatattgg cgggtaaact aagtcgctgt
     3721 atgtgtttgt ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4201 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4261 ctctgctgaa gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa
     4321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4501 attaaaaatg aagttttaaa tcaatctaaa gtatatatgt gtaacattgg tctagtgatt
     4561 atttgccgac taccttggtg atctcgcctt tcacgtagtg aacaaattct tccaactgat
     4621 ctgcgcgcga ggccaagcga tcttcttgtc caagataagc ctgcctagct tcaagtatga
     4681 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     4741 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     4801 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     4861 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     4921 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     4981 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     5041 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     5101 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     5161 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     5221 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     5281 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     5341 ccctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     5401 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     5461 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     5521 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     5581 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcgatcccca
     5641 tccaacagcc cgccgtcgag cgggcttttt tatccccgga agcctgtgga tagagggtag
     5701 ttatccacgt gaaaccgcta atgccccgca aagccttgat tcacggggct ttccggcccg
     5761 ctccaaaaac tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct aatcggagta
     5821 cgtgaaatcg ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta
     5881 atagcccttt cagatcaaca gcttgcaaac acccctcgct ccggcaagta gttacagcaa
     5941 gtagtatgtt caattagctt ttcaattatg aatatatata tcaattattg gtcgcccttg
     6001 gcttgtggac aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc
     6061 caccgtcgag cgcctttgcc cacaacccgg cggccggccg caacagatcg ttttataaat
     6121 tttttttttt gaaaaagaaa aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc
     6181 gatccccgga attagatctt ggcaggatat attgtggtgt aacgttctcg aactaggctc
     6241 ggacgaagta agcggccgca accttaatta aacacggcgc gccaacagcg atcgcaccag
     6301 gtaccacctc cgatccattc ggtccccaga ttagcctttt caatttcaga aagaatgcta
     6361 acccacagat ggttagagag gcttacgcag cacgtctcat caagacgatc tacccgagca
     6421 ataatctcca ggaaatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca
     6481 ggactaactg catcaagaac acagagaaag atatatttct caagatcaga agtactattc
     6541 cagtatggac gattcaaggc ttgcttcaca aaccaaggca agtaatagag attggagtct
     6601 ctaaaaaggt agttcccac
//
