LOCUS       sequence-239408-        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"
     promoter        45..390
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     333..353
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             441..827
                     /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             882..902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1791..2432
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    2538..2562
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2653..3241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2733..2752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3412..4200)
                     /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      4491..4926
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5053..5075
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac ttttcaacaa
       61 agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca ctttattgtg
      121 aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggcc
      181 atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
      241 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
      301 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
      361 taaggaagtt catttcattt ggagagaaca cgggggacga gctaacagta ttcagtcgac
      421 tggtaccaac aggctccatg gcttctaatt ttactcaatt tgttttggtt gataatggag
      481 gaactggaga tgttactgtt gctccttcta attttgctaa tggagttgct gaatggattt
      541 cttctaattc aagatctcag gcttataagg ttacttgttc tgttagacaa tcttctgctc
      601 aaaagagaaa gtatactatt aaggttgaag ttcctaaggt tgctactcaa actgttggag
      661 gagttgaatt gcctgttgct gcttggagat cttatttgaa tatggaattg actattccta
      721 tttttgctac taattctgat tgtgaattga ttgttaaggc tatgcaagga ttgttgaagg
      781 atggaaatcc tattccttct gctattgctg ctaattctgg aatttattct gctggaggag
      841 gaggatctgg aggaggagga tctggaggag gaggatctgg acctaagaag aagagaaagg
      901 ttgctgctgc tggatcttca ggtatggctg gtgagaatgt gccaatccct tgtgtcaatg
      961 gggtagatgg agaaccttgt cctgaagatt acaagtacat atcggaaaat tgcgagacaa
     1021 gcacgatgaa tatcgatcgg aacattactc atctccaaca ctgtacatgt gttgatgact
     1081 gctcatcttc aaactgtctt tgcggtcaat tgtcaatcag atgttggtac gataaagacg
     1141 gaagattgct tcaggagttt aacaaaatcg aaccacctct catctttgag tgtaatcaag
     1201 cctgcagttg ttggagatct tgcaagaatc gagttgtcca aagtggcatc aaagttcgtt
     1261 tacagctcta tcgtactgct aaaatgggat ggggtgtaag agcacttcag accattccac
     1321 aaggtacctt catttgcgag tatgtgggag agctaatttc cgacgctgaa gcagacgtta
     1381 gagaggatga tagctacttg tttgacctcg ataacaagga tggcgaagtc tactgcatag
     1441 atgctcgata ttatgggaac atttctaggt tcataaacca tctgtgtgat cccaatatca
     1501 ttccagtgag agttttcatg ttacaccagg atttacggtt tcctaggatt gccttctttt
     1561 ctagcagaga tatacgtaca ggagaggaat tggggtttga ctatggtgac agattctggg
     1621 atattaagtc caagtatttc acttgtcagt gtggatctga gaaatgcaag catagtgctg
     1681 aagcgattgc gttggaacaa tctaggctag caaggcttga tccgcatcct gaactgcttc
     1741 ccgatctttc gtcactacca ccgataaaca cgactggtta gtaactgcaa agctttcgtt
     1801 cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca
     1861 ccagaatcct actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt
     1921 tgttgtgctt gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg
     1981 aaatggatgg agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat
     2041 tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac
     2101 attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag
     2161 gagtaaaaca cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca
     2221 gacctagaaa agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt
     2281 tatgaacttt cctttatgta attttccaga atccttgtca gattctaatc attgctttat
     2341 aattatagtt atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta
     2401 tgacttgcca attgattgac aacatgcatc aatcgactat gagtaggcaa gatgttctgt
     2461 aggaccttga gaccgaaaag gtggtctcag tattctagac caagcatgca cagctggcgt
     2521 tctcagccag atccccttga caggatatat tggcgggtaa actaagtcgc tgtatgtgtt
     2581 tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3121 gaagccagtt accttcggaa gaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3361 atgaagtttt aaatcaatct aaagtatata tgtgtaacat tggtctagtg attatttgcc
     3421 gactaccttg gtgatctcgc ctttcacgta gtgaacaaat tcttccaact gatctgcgcg
     3481 cgaggccaag cgatcttctt gtccaagata agcctgccta gcttcaagta tgacgggctg
     3541 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     3601 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     3661 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     3721 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     3781 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     3841 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     3901 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     3961 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     4021 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     4081 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     4141 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     4201 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     4261 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     4321 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     4381 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     4441 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcgatcc ccatccaaca
     4501 gcccgccgtc gagcgggctt ttttatcccc ggaagcctgt ggatagaggg tagttatcca
     4561 cgtgaaaccg ctaatgcccc gcaaagcctt gattcacggg gctttccggc ccgctccaaa
     4621 aactatccac gtgaaatcgc taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa
     4681 tcgctaataa ggtcacgtga aatcgctaat caaaaaggca cgtgagaacg ctaatagccc
     4741 tttcagatca acagcttgca aacacccctc gctccggcaa gtagttacag caagtagtat
     4801 gttcaattag cttttcaatt atgaatatat atatcaatta ttggtcgccc ttggcttgtg
     4861 gacaatgcgc tacgcgcacc ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc
     4921 gagcgccttt gcccacaacc cggcggccgg ccgcaacaga tcgttttata aatttttttt
     4981 tttgaaaaag aaaaagcccg aaaggcggca acctctcggg cttctggatt tccgatcccc
     5041 ggaattagat cttggcagga tatattgtgg tgtaacgttc tcgaactagg ctcggacgaa
     5101 gtaagcggcc gcaaccttaa ttaaacacgg cgcgccaaca gcgatcgcac caggtaccac
     5161 ctccgatcca ttcggtcccc agattagcct tttcaatttc agaaagaatg ctaacccaca
     5221 gatggttaga gaggcttacg cagcacgtct catcaagacg atctacccga gcaataatct
     5281 ccaggaaatc aaataccttc ccaagaaggt taaagatgca gtcaaaagat tcaggactaa
     5341 ctgcatcaag aacacagaga aagatatatt tctcaagatc agaagtacta ttccagtatg
     5401 gacgattcaa ggcttgcttc acaaaccaag gcaagtaata gagattggag tctctaaaaa
     5461 ggtagttccc actgaatcaa aggccatgga gtcaaagatt caaatagagg acctaacaga
     5521 actcgccgta aagactggcg aacagttcat acagagtctc ttacgactca atgacaagaa
     5581 gaaaatcttc gtcaacatgg tggagcacga cacacttgtc tac
//
