LOCUS       sequence-234670-        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"
     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      2841..3482
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    3588..3612
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3703..4291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3783..3802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4462..5250)
                     /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(5541..5569,1..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 gacatctccg
     2041 gagggctaga atttaagggt atccccgcca ctaatcgtgt tgatgattca ccagtttcac
     2101 caacatctgg tttcacatac atcaaatctt tgattattga gcctaatgtc ataattccaa
     2161 agagttcaac tgggtgtaac tgccgaggca gctgcactga ctcaaagaaa tgtgcatgtg
     2221 ctaagcttaa tgggggtaac tttccatatg ttgaccttaa tgatggcaga ttaattgagt
     2281 ctcgagatgt tgtatttgaa tgtggtcctc actgtgggtg tgggccaaaa tgtgtcaacc
     2341 gaacttctca gaagcgtcta agattcaatc ttgaggtttt ccgctctgca aagaagggtt
     2401 gggcagttag atcatgggag tacataccag ctggttcacc agtatgtgag tacataggag
     2461 ttgtcaggag aactgctgat gtggatacta tctctgacaa tgaatacata tttgagattg
     2521 actgccaaca gacaatgcaa ggtcttggtg gaagacagag aagactaaga gatgttgctg
     2581 taccaatgaa taatggagtc agtcagagca gtgaagatga gaatgcgcca gagttctgca
     2641 ttgatgctgg ttcaacagga aactttgcta ggtttataaa tcacagttgt gaaccaaacc
     2701 tatttgttca gtgcgtcctg agttctcacc aggatataag gcttgcccgt gtggttcttt
     2761 tcgcagctga caacatttcc ccaatgcagg agctcactta cgactatgga tatgcgcttg
     2821 atagcgttca ttaactgcaa agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa
     2881 gttcaatgca tcagtttcat tgcgcacaca ccagaatcct actgagtttg agtattatgg
     2941 cattgggaaa actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta
     3001 ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata
     3061 tggtcctttt gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt
     3121 tgaatttgaa attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc
     3181 gtggcctcta atgaccgaag ttaatatgag gagtaaaaca cttgtagttg taccattatg
     3241 cttattcact aggcaacaaa tatattttca gacctagaaa agctgcaaat gttactgaat
     3301 acaagtatgt cctcttgtgt tttagacatt tatgaacttt cctttatgta attttccaga
     3361 atccttgtca gattctaatc attgctttat aattatagtt atactcatgg atttgtagtt
     3421 gagtatgaaa atatttttta atgcatttta tgacttgcca attgattgac aacatgcatc
     3481 aatcgactat gagtaggcaa gatgttctgt aggaccttga gaccgaaaag gtggtctcag
     3541 tattctagac caagcatgca cagctggcgt tctcagccag atccccttga caggatatat
     3601 tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca
     3661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa gaagagttgg
     4201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4441 tgtgtaacat tggtctagtg attatttgcc gactaccttg gtgatctcgc ctttcacgta
     4501 gtgaacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt gtccaagata
     4561 agcctgccta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     4621 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     4681 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
     4741 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     4801 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     4861 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     4921 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     4981 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     5041 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     5101 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     5161 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     5221 cgatacttcg gcgatcaccg cttccctcat aacacccctt gtattactgt ttatgtaagc
     5281 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     5341 ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
     5401 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
     5461 ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga
     5521 tggggcgatt caggcgatcc ccatccaaca gcccgccgtc gagcgggct
//
