LOCUS       sequence-239407-        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"
     promoter        68..413
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     356..376
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             464..850
                     /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             905..925
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      998..1639
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    1745..1769
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1860..2448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1940..1959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2619..3407)
                     /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      3698..4133
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4260..4282
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa
       61 gggcaattga gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc
      121 cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc
      181 atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag
      241 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
      301 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
      361 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagaga acacggggga
      421 cgagctaaca gtattcagtc gactggtacc aacaggctcc atggcttcta attttactca
      481 atttgttttg gttgataatg gaggaactgg agatgttact gttgctcctt ctaattttgc
      541 taatggagtt gctgaatgga tttcttctaa ttcaagatct caggcttata aggttacttg
      601 ttctgttaga caatcttctg ctcaaaagag aaagtatact attaaggttg aagttcctaa
      661 ggttgctact caaactgttg gaggagttga attgcctgtt gctgcttgga gatcttattt
      721 gaatatggaa ttgactattc ctatttttgc tactaattct gattgtgaat tgattgttaa
      781 ggctatgcaa ggattgttga aggatggaaa tcctattcct tctgctattg ctgctaattc
      841 tggaatttat tctgctggag gaggaggatc tggaggagga ggatctggag gaggaggatc
      901 tggacctaag aagaagagaa aggttgctgc tgctggatct tcaggtatgc ttgatctcga
      961 cttggagctt agactcggat tcgcttaata gctgcaaagc tttcgttcgt atcatcggtt
     1021 tcgacaacgt tcgtcaagtt caatgcatca gtttcattgc gcacacacca gaatcctact
     1081 gagtttgagt attatggcat tgggaaaact gtttttcttg taccatttgt tgtgcttgta
     1141 atttactgtg ttttttattc ggttttcgct atcgaactgt gaaatggaaa tggatggaga
     1201 agagttaatg aatgatatgg tccttttgtt cattctcaaa ttaatattat ttgttttttc
     1261 tcttatttgt tgtgtgttga atttgaaatt ataagagata tgcaaacatt ttgttttgag
     1321 taaaaatgtg tcaaatcgtg gcctctaatg accgaagtta atatgaggag taaaacactt
     1381 gtagttgtac cattatgctt attcactagg caacaaatat attttcagac ctagaaaagc
     1441 tgcaaatgtt actgaataca agtatgtcct cttgtgtttt agacatttat gaactttcct
     1501 ttatgtaatt ttccagaatc cttgtcagat tctaatcatt gctttataat tatagttata
     1561 ctcatggatt tgtagttgag tatgaaaata ttttttaatg cattttatga cttgccaatt
     1621 gattgacaac atgcatcaat cgactatgag taggcaagat gttctgtagg accttgagac
     1681 cgaaaaggtg gtctcagtat tctagaccaa gcatgcacag ctggcgttct cagccagatc
     1741 cccttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc
     1801 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1861 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1981 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2041 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2101 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2161 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2221 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2281 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2341 ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2401 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2461 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2521 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2581 tcaatctaaa gtatatatgt gtaacattgg tctagtgatt atttgccgac taccttggtg
     2641 atctcgcctt tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga
     2701 tcttcttgtc caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc
     2761 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     2821 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     2881 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     2941 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     3001 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     3061 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     3121 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     3181 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     3241 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
     3301 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     3361 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataac accccttgta
     3421 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3481 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     3541 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     3601 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     3661 actttctggc tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag
     3721 cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta
     3781 atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg
     3841 aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt
     3901 cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca
     3961 gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt
     4021 ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac
     4081 gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc
     4141 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
     4201 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagatctt
     4261 ggcaggatat attgtggtgt aacgttctcg aactaggctc ggacgaagta agcggccgca
     4321 accttaatta aacacggcgc gccaacagcg atcgcaccag gtaccacctc cgatccattc
     4381 ggtccccaga ttagcctttt caatttcaga aagaatgcta acccacagat ggttagagag
     4441 gcttacgcag cacgtctcat caagacgatc tacccgagca ataatctcca ggaaatcaaa
     4501 taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg catcaagaac
     4561 acagagaaag atatatttct caagatcaga agtactattc cagtatggac gattcaaggc
     4621 ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt agttcccact
     4681 gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact cgccgtaaag
     4741 actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa aatcttcgtc
     4801 aacatgg
//
