LOCUS       sequence-239409-        5569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector containing 35S:MS2-KYP: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 5569)
  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 5569)
  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: pJL042
FEATURES             Location/Qualifiers
     source          1..5569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        167..512
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     455..475
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             563..949
                     /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             1004..1024
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1859..2500
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    2606..2630
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2721..3309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2801..2820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3480..4268)
                     /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      4559..4994
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5121..5143
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
       61 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
      121 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac
      181 aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg
      241 tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg
      301 ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
      361 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
      421 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
      481 tataaggaag ttcatttcat ttggagagaa cacgggggac gagctaacag tattcagtcg
      541 actggtacca acaggctcca tggcttctaa ttttactcaa tttgttttgg ttgataatgg
      601 aggaactgga gatgttactg ttgctccttc taattttgct aatggagttg ctgaatggat
      661 ttcttctaat tcaagatctc aggcttataa ggttacttgt tctgttagac aatcttctgc
      721 tcaaaagaga aagtatacta ttaaggttga agttcctaag gttgctactc aaactgttgg
      781 aggagttgaa ttgcctgttg ctgcttggag atcttatttg aatatggaat tgactattcc
      841 tatttttgct actaattctg attgtgaatt gattgttaag gctatgcaag gattgttgaa
      901 ggatggaaat cctattcctt ctgctattgc tgctaattct ggaatttatt ctgctggagg
      961 aggaggatct ggaggaggag gatctggagg aggaggatct ggacctaaga agaagagaaa
     1021 ggttgctgct gctggatctt caggtatgga catctccgga gggctagaat ttaagggtat
     1081 ccccgccact aatcgtgttg atgattcacc agtttcacca acatctggtt tcacatacat
     1141 caaatctttg attattgagc ctaatgtcat aattccaaag agttcaactg ggtgtaactg
     1201 ccgaggcagc tgcactgact caaagaaatg tgcatgtgct aagcttaatg ggggtaactt
     1261 tccatatgtt gaccttaatg atggcagatt aattgagtct cgagatgttg tatttgaatg
     1321 tggtcctcac tgtgggtgtg ggccaaaatg tgtcaaccga acttctcaga agcgtctaag
     1381 attcaatctt gaggttttcc gctctgcaaa gaagggttgg gcagttagat catgggagta
     1441 cataccagct ggttcaccag tatgtgagta cataggagtt gtcaggagaa ctgctgatgt
     1501 ggatactatc tctgacaatg aatacatatt tgagattgac tgccaacaga caatgcaagg
     1561 tcttggtgga agacagagaa gactaagaga tgttgctgta ccaatgaata atggagtcag
     1621 tcagagcagt gaagatgaga atgcgccaga gttctgcatt gatgctggtt caacaggaaa
     1681 ctttgctagg tttataaatc acagttgtga accaaaccta tttgttcagt gcgtcctgag
     1741 ttctcaccag gatataaggc ttgcccgtgt ggttcttttc gcagctgaca acatttcccc
     1801 aatgcaggag ctcacttacg actatggata tgcgcttgat agcgttcatt aactgcaaag
     1861 ctttcgttcg tatcatcggt ttcgacaacg ttcgtcaagt tcaatgcatc agtttcattg
     1921 cgcacacacc agaatcctac tgagtttgag tattatggca ttgggaaaac tgtttttctt
     1981 gtaccatttg ttgtgcttgt aatttactgt gttttttatt cggttttcgc tatcgaactg
     2041 tgaaatggaa atggatggag aagagttaat gaatgatatg gtccttttgt tcattctcaa
     2101 attaatatta tttgtttttt ctcttatttg ttgtgtgttg aatttgaaat tataagagat
     2161 atgcaaacat tttgttttga gtaaaaatgt gtcaaatcgt ggcctctaat gaccgaagtt
     2221 aatatgagga gtaaaacact tgtagttgta ccattatgct tattcactag gcaacaaata
     2281 tattttcaga cctagaaaag ctgcaaatgt tactgaatac aagtatgtcc tcttgtgttt
     2341 tagacattta tgaactttcc tttatgtaat tttccagaat ccttgtcaga ttctaatcat
     2401 tgctttataa ttatagttat actcatggat ttgtagttga gtatgaaaat attttttaat
     2461 gcattttatg acttgccaat tgattgacaa catgcatcaa tcgactatga gtaggcaaga
     2521 tgttctgtag gaccttgaga ccgaaaaggt ggtctcagta ttctagacca agcatgcaca
     2581 gctggcgttc tcagccagat ccccttgaca ggatatattg gcgggtaaac taagtcgctg
     2641 tatgtgtttg tttgagatct catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3181 gctctgctga agccagttac cttcggaaga agagttggta gctcttgatc cggcaaacaa
     3241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3421 aattaaaaat gaagttttaa atcaatctaa agtatatatg tgtaacattg gtctagtgat
     3481 tatttgccga ctaccttggt gatctcgcct ttcacgtagt gaacaaattc ttccaactga
     3541 tctgcgcgcg aggccaagcg atcttcttgt ccaagataag cctgcctagc ttcaagtatg
     3601 acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc
     3661 gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc
     3721 tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca
     3781 aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca
     3841 acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc
     3901 tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta
     3961 gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg
     4021 agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc
     4081 cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc
     4141 ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga
     4201 tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct
     4261 tccctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     4321 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt
     4381 tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc cgacaacgca
     4441 gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta caacaaagct
     4501 ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca ggcgatcccc
     4561 atccaacagc ccgccgtcga gcgggctttt ttatccccgg aagcctgtgg atagagggta
     4621 gttatccacg tgaaaccgct aatgccccgc aaagccttga ttcacggggc tttccggccc
     4681 gctccaaaaa ctatccacgt gaaatcgcta atcagggtac gtgaaatcgc taatcggagt
     4741 acgtgaaatc gctaataagg tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct
     4801 aatagccctt tcagatcaac agcttgcaaa cacccctcgc tccggcaagt agttacagca
     4861 agtagtatgt tcaattagct tttcaattat gaatatatat atcaattatt ggtcgccctt
     4921 ggcttgtgga caatgcgcta cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc
     4981 ccaccgtcga gcgcctttgc ccacaacccg gcggccggcc gcaacagatc gttttataaa
     5041 tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct tctggatttc
     5101 cgatccccgg aattagatct tggcaggata tattgtggtg taacgttctc gaactaggct
     5161 cggacgaagt aagcggccgc aaccttaatt aaacacggcg cgccaacagc gatcgcacca
     5221 ggtaccacct ccgatccatt cggtccccag attagccttt tcaatttcag aaagaatgct
     5281 aacccacaga tggttagaga ggcttacgca gcacgtctca tcaagacgat ctacccgagc
     5341 aataatctcc aggaaatcaa ataccttccc aagaaggtta aagatgcagt caaaagattc
     5401 aggactaact gcatcaagaa cacagagaaa gatatatttc tcaagatcag aagtactatt
     5461 ccagtatgga cgattcaagg cttgcttcac aaaccaaggc aagtaataga gattggagtc
     5521 tctaaaaagg tagttcccac tgaatcaaag gccatggagt caaagattc
//
