LOCUS       sequence-234668-        4807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector containing 35S:MS2-SRDX: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 4807)
  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 4807)
  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: pJL009
FEATURES             Location/Qualifiers
     source          1..4807
                     /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      2079..2720
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    2826..2850
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2941..3529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3021..3040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3700..4488)
                     /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(4779..4807,1..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 cttgatctcg
     2041 acttggagct tagactcgga ttcgcttaat agctgcaaag ctttcgttcg tatcatcggt
     2101 ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg cgcacacacc agaatcctac
     2161 tgagtttgag tattatggca ttgggaaaac tgtttttctt gtaccatttg ttgtgcttgt
     2221 aatttactgt gttttttatt cggttttcgc tatcgaactg tgaaatggaa atggatggag
     2281 aagagttaat gaatgatatg gtccttttgt tcattctcaa attaatatta tttgtttttt
     2341 ctcttatttg ttgtgtgttg aatttgaaat tataagagat atgcaaacat tttgttttga
     2401 gtaaaaatgt gtcaaatcgt ggcctctaat gaccgaagtt aatatgagga gtaaaacact
     2461 tgtagttgta ccattatgct tattcactag gcaacaaata tattttcaga cctagaaaag
     2521 ctgcaaatgt tactgaatac aagtatgtcc tcttgtgttt tagacattta tgaactttcc
     2581 tttatgtaat tttccagaat ccttgtcaga ttctaatcat tgctttataa ttatagttat
     2641 actcatggat ttgtagttga gtatgaaaat attttttaat gcattttatg acttgccaat
     2701 tgattgacaa catgcatcaa tcgactatga gtaggcaaga tgttctgtag gaccttgaga
     2761 ccgaaaaggt ggtctcagta ttctagacca agcatgcaca gctggcgttc tcagccagat
     2821 ccccttgaca ggatatattg gcgggtaaac taagtcgctg tatgtgtttg tttgagatct
     2881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3421 cttcggaaga agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3661 atcaatctaa agtatatatg tgtaacattg gtctagtgat tatttgccga ctaccttggt
     3721 gatctcgcct ttcacgtagt gaacaaattc ttccaactga tctgcgcgcg aggccaagcg
     3781 atcttcttgt ccaagataag cctgcctagc ttcaagtatg acgggctgat actgggccgg
     3841 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     3901 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     3961 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     4021 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     4081 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     4141 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     4201 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     4261 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     4321 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     4381 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     4441 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcataa caccccttgt
     4501 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     4561 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc
     4621 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     4681 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct
     4741 cactttctgg ctggatgatg gggcgattca ggcgatcccc atccaacagc ccgccgtcga
     4801 gcgggct
//
