LOCUS       sequence-234669-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector containing 35S:MS2-G9a: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 5623)
  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 5623)
  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: pJL036
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    534..556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     promoter        1149..1494
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1437..1457
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1545..1931
                     /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             1986..2006
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      2895..3536
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    3642..3666
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3757..4345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3837..3856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4516..5304)
                     /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      join(5595..5623,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
      421 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
      481 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg
      541 atatattgtg gtgtaacgtt ctcgaactag gctcggacga agtaagcggc cgcaacctta
      601 attaaacacg gcgcgccaac agcgatcgca ccaggtacca cctccgatcc attcggtccc
      661 cagattagcc ttttcaattt cagaaagaat gctaacccac agatggttag agaggcttac
      721 gcagcacgtc tcatcaagac gatctacccg agcaataatc tccaggaaat caaatacctt
      781 cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta actgcatcaa gaacacagag
      841 aaagatatat ttctcaagat cagaagtact attccagtat ggacgattca aggcttgctt
      901 cacaaaccaa ggcaagtaat agagattgga gtctctaaaa aggtagttcc cactgaatca
      961 aaggccatgg agtcaaagat tcaaatagag gacctaacag aactcgccgt aaagactggc
     1021 gaacagttca tacagagtct cttacgactc aatgacaaga agaaaatctt cgtcaacatg
     1081 gtggagcacg acacacttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa
     1141 agggcaattg agacttttca acaaagggta atatccggaa acctcctcgg attccattgc
     1201 ccagctatct gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc
     1261 catcattgcg ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa
     1321 gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca
     1381 aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat
     1441 ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag aacacggggg
     1501 acgagctaac agtattcagt cgactggtac caacaggctc catggcttct aattttactc
     1561 aatttgtttt ggttgataat ggaggaactg gagatgttac tgttgctcct tctaattttg
     1621 ctaatggagt tgctgaatgg atttcttcta attcaagatc tcaggcttat aaggttactt
     1681 gttctgttag acaatcttct gctcaaaaga gaaagtatac tattaaggtt gaagttccta
     1741 aggttgctac tcaaactgtt ggaggagttg aattgcctgt tgctgcttgg agatcttatt
     1801 tgaatatgga attgactatt cctatttttg ctactaattc tgattgtgaa ttgattgtta
     1861 aggctatgca aggattgttg aaggatggaa atcctattcc ttctgctatt gctgctaatt
     1921 ctggaattta ttctgctgga ggaggaggat ctggaggagg aggatctgga ggaggaggat
     1981 ctggacctaa gaagaagaga aaggttgctg ctgctggatc ttcaggtatg gctggtgaga
     2041 atgtgccaat cccttgtgtc aatggggtag atggagaacc ttgtcctgaa gattacaagt
     2101 acatatcgga aaattgcgag acaagcacga tgaatatcga tcggaacatt actcatctcc
     2161 aacactgtac atgtgttgat gactgctcat cttcaaactg tctttgcggt caattgtcaa
     2221 tcagatgttg gtacgataaa gacggaagat tgcttcagga gtttaacaaa atcgaaccac
     2281 ctctcatctt tgagtgtaat caagcctgca gttgttggag atcttgcaag aatcgagttg
     2341 tccaaagtgg catcaaagtt cgtttacagc tctatcgtac tgctaaaatg ggatggggtg
     2401 taagagcact tcagaccatt ccacaaggta ccttcatttg cgagtatgtg ggagagctaa
     2461 tttccgacgc tgaagcagac gttagagagg atgatagcta cttgtttgac ctcgataaca
     2521 aggatggcga agtctactgc atagatgctc gatattatgg gaacatttct aggttcataa
     2581 accatctgtg tgatcccaat atcattccag tgagagtttt catgttacac caggatttac
     2641 ggtttcctag gattgccttc ttttctagca gagatatacg tacaggagag gaattggggt
     2701 ttgactatgg tgacagattc tgggatatta agtccaagta tttcacttgt cagtgtggat
     2761 ctgagaaatg caagcatagt gctgaagcga ttgcgttgga acaatctagg ctagcaaggc
     2821 ttgatccgca tcctgaactg cttcccgatc tttcgtcact accaccgata aacacgactg
     2881 gttagtaact gcaaagcttt cgttcgtatc atcggtttcg acaacgttcg tcaagttcaa
     2941 tgcatcagtt tcattgcgca cacaccagaa tcctactgag tttgagtatt atggcattgg
     3001 gaaaactgtt tttcttgtac catttgttgt gcttgtaatt tactgtgttt tttattcggt
     3061 tttcgctatc gaactgtgaa atggaaatgg atggagaaga gttaatgaat gatatggtcc
     3121 ttttgttcat tctcaaatta atattatttg ttttttctct tatttgttgt gtgttgaatt
     3181 tgaaattata agagatatgc aaacattttg ttttgagtaa aaatgtgtca aatcgtggcc
     3241 tctaatgacc gaagttaata tgaggagtaa aacacttgta gttgtaccat tatgcttatt
     3301 cactaggcaa caaatatatt ttcagaccta gaaaagctgc aaatgttact gaatacaagt
     3361 atgtcctctt gtgttttaga catttatgaa ctttccttta tgtaattttc cagaatcctt
     3421 gtcagattct aatcattgct ttataattat agttatactc atggatttgt agttgagtat
     3481 gaaaatattt tttaatgcat tttatgactt gccaattgat tgacaacatg catcaatcga
     3541 ctatgagtag gcaagatgtt ctgtaggacc ttgagaccga aaaggtggtc tcagtattct
     3601 agaccaagca tgcacagctg gcgttctcag ccagatcccc ttgacaggat atattggcgg
     3661 gtaaactaag tcgctgtatg tgtttgtttg agatctcatg tgagcaaaag gccagcaaaa
     3721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaagaagag ttggtagctc
     4261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgtgta
     4501 acattggtct agtgattatt tgccgactac cttggtgatc tcgcctttca cgtagtgaac
     4561 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttgtccaa gataagcctg
     4621 cctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     4681 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     4741 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     4801 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     4861 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     4921 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     4981 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     5041 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     5101 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     5161 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     5221 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     5281 ttcggcgatc accgcttccc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     5341 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     5401 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     5461 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     5521 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     5581 gattcaggcg atccccatcc aacagcccgc cgtcgagcgg gct
//
