LOCUS       sequence-234667-        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"
     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      3891..4532
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    4638..4662
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4753..5341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4833..4852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5512..6300)
                     /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(6591..6619,1..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 ttcaggtatt tttaagccag
     2041 aggagcttag acaagcactt atgcctactc ttgaggctct ttacagacaa gatcctgaat
     2101 ctcttccatt cagacagcct gtggaccctc agcttcttgg tattcctgac tacttcgata
     2161 ttgtgaagtc tcctatggat ctttctacaa ttaaaagaaa acttgacaca ggacaatacc
     2221 aagaaccttg gcaatacgtt gatgacatct ggcttatgtt taacaatgct tggctttaca
     2281 acagaaagac ttctagagtt tacaaatatt gctctaaact ttctgaagtg ttcgaacaag
     2341 aaattgatcc agtgatgcaa tctcttggtt actgttgcgg tagaaaactt gagttctctc
     2401 ctcaaactct ttgctgctat ggaaagcaac tttgtactat tcctagagat gcaacatact
     2461 attcttatca aaacagatac catttctgtg aaaaatgctt taacgaaatc cagggtgaat
     2521 ctgtttctct tggagacgac ccttctcagc ctcagacaac aatcaataaa gagcagttct
     2581 ctaagagaaa gaacgatact cttgaccctg agcttttcgt ggagtgtact gagtgcggta
     2641 gaaagatgca tcagatttgt gttcttcatc acgagatcat ttggccagct ggttttgttt
     2701 gcgatggttg tcttaagaag tctgcaagaa ctagaaaaga gaataagttc tctgcaaaga
     2761 gacttccttc tactagactt ggaactttcc ttgaaaacag agtgaacgat ttccttagaa
     2821 gacagaatca tcctgagtct ggagaggtga cagtgagagt ggttcatgca tctgataaaa
     2881 ctgttgaggt taaacctgga atgaaagcta gattcgtgga ctctggagag atggcagaat
     2941 cttttccata cagaacaaaa gctctttttg cattcgaaga gattgatggt gttgatcttt
     3001 gtttcttcgg tatgcacgtg caagagtatg gatctgactg ccctccacca aatcaaagaa
     3061 gagtttacat ttcttatctt gattctgttc acttttttag accaaaatgc cttagaacag
     3121 cagtgtacca cgagattctt attggttatc ttgagtatgt gaaaaaactt ggttacacaa
     3181 ctggacatat ttgggcttgc ccaccttctg aaggtgatga ttatattttt cattgccacc
     3241 ctccagacca aaagattcct aaacctaaaa gacttcaaga gtggtacaaa aagatgcttg
     3301 ataaagcagt gtctgagaga attgtgcacg actacaagga catttttaaa caggctacag
     3361 aggatagact tacttctgca aaagagcttc cttattttga aggagatttt tggcctaatg
     3421 ttcttgagga gtctattaag gaacttgagc aggaggaaga agagagaaaa agagaagaga
     3481 acacatctaa tgagtctaca gacgttacta aaggtgattc taaaaacgct aaaaagaaaa
     3541 acaataaaaa aacttctaaa aataaatctt ctctttctag aggtaacaaa aaaaaacctg
     3601 gaatgccaaa cgtttctaat gacctttctc aaaaacttta cgcaacaatg gaaaaacata
     3661 aagaagtttt cttcgtgatc agacttattg ctggacctgc tgctaattct cttcctccaa
     3721 tcgttgaccc agaccctctt attccttgtg atcttatgga tggaagagac gcatttctta
     3781 ctcttgcaag agacaaacat cttgaatttt cttctcttag aagagcacag tggtctacaa
     3841 tgtgtatgct tgttgagctt cacacacaat ctcaagacta gtaactgcaa agctttcgtt
     3901 cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca
     3961 ccagaatcct actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt
     4021 tgttgtgctt gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg
     4081 aaatggatgg agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat
     4141 tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac
     4201 attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag
     4261 gagtaaaaca cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca
     4321 gacctagaaa agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt
     4381 tatgaacttt cctttatgta attttccaga atccttgtca gattctaatc attgctttat
     4441 aattatagtt atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta
     4501 tgacttgcca attgattgac aacatgcatc aatcgactat gagtaggcaa gatgttctgt
     4561 aggaccttga gaccgaaaag gtggtctcag tattctagac caagcatgca cagctggcgt
     4621 tctcagccag atccccttga caggatatat tggcgggtaa actaagtcgc tgtatgtgtt
     4681 tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5221 gaagccagtt accttcggaa gaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5461 atgaagtttt aaatcaatct aaagtatata tgtgtaacat tggtctagtg attatttgcc
     5521 gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact gatctgcgcg
     5581 cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta tgacgggctg
     5641 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     5701 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     5761 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     5821 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     5881 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     5941 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     6001 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     6061 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     6121 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     6181 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     6241 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     6301 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     6361 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     6421 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     6481 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     6541 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcgatcc ccatccaaca
     6601 gcccgccgtc gagcgggct
//
