LOCUS       sequence-239404-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector containing pSUC2:MS2-VP64: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 6199)
  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 6199)
  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: pMH205
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1948..2334
                     /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             2389..2409
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2440..2589
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     terminator      2608..3249
                     /label=E9 terminator
                     /note="terminator and polyadenylation signal from the pea 
                     rbcS-E9 gene"
     misc_feature    3355..3379
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3470..4058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3550..3569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4229..5017)
                     /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      5308..5743
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5870..5892
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 gtctataaga attcggaaac atatagtatc ctttgaaaac ggagaaacaa ataacaatgt
       61 ggacaaacta gatataattt caacacaaga ctatgggaat gattttaccc actaattata
      121 atccgatcac aaggtttcaa cgaactagtt ttccagatat caaccaaatt tactttggaa
      181 ttaaactaac ttaaaactaa ttggttgttc gtaaatggtg cttttttttt ttgcggatgt
      241 tagtaaaggg ttttatgtat tttatattat tagttatctg ttttcagtgt tatgttgtct
      301 catccataaa gtttatatgt tttttctttg ctctataact tatatatata tatgagttta
      361 cagttatatt tatacatttc agatacttga tcggcatttt ttttggtaaa aaatatatgc
      421 atgaaaaact caagtgtttc ttttttaagg aatttttaaa tggtgattat atgaatataa
      481 tcatatgtat atccgtatat atatgtagcc agatagttaa ttatttgggg gatatttgaa
      541 ttattaatgt tataatattc tttcttttga ctcgtctggt taaattaaag aacaaaaaaa
      601 acacatactt ttactgtttt aaaaggttaa attaacataa tttattgatt acaagtgtca
      661 agtccatgac attgcatgta ggttcgagac ttcagagata acggaagaga tcgataattg
      721 tgatcgtaac atccagatat gtatgtttaa ttttcattta gatgtggatc agagaagata
      781 agtcaaactg tcttcataat ttaagacaac ctcttttaat attttcccaa aacatgtttt
      841 atgtaactac tttgcttatg tgattgcctg aggatactat tattctctgt ctttattctc
      901 ttcacaccac atttaaatag tttaagagca tagaaattaa ttattttcaa aaaggtgatt
      961 atatgcatgc aaaatagcac accatttatg tttatatttt caaattattt aatacatttc
     1021 aatatttcat aagtgtgatt ttttttttta ttgtcaattt cataagtgtg atttgtcatt
     1081 tgtattaaac aattgtatcg cgcagtacaa ataaacagtg ggagaggtga aaatgcagtt
     1141 ataaaactgt ccaataattt actaacacat ttaaatatct aaaaagagtg tttcaaaaaa
     1201 aattcttttg aaataagaaa agtgatagat atttttacgc tttcgtctga aaataaaaca
     1261 ataatagttt attagaaaaa tgttatcacc gaaaattatt ctagtgccac tcgctcggat
     1321 cgaaattcga aagttatatt ctttctcttt acctaatata aaaatcacaa gaaaaatcaa
     1381 tccgaatata tctatcaaca tagtatatgc ccttacatat tgtttctgac ttttctctat
     1441 ccgaatttct cgcttcatgg ttttttttta acatattctc atttaatttt cattactatt
     1501 atataactaa aagatggaaa taaaataaag tgtctttgag aatcgaacgt ccatatcagt
     1561 aagatagttt gtgtgaaggt aaaatctaaa agatttaagt tccaaaaaca gaaaataata
     1621 tattacgcta aaaaagaaga aaataattaa atacaaaaca gaaaaaaata atatacgaca
     1681 gacacgtgtc acgaagatac cctacgctat agacacagct ctgttttctc ttttctatgc
     1741 ctcaaggctc tcttaacttc actgtctcct cttcggataa tcctatcctt ctcttcctat
     1801 aaatacctct ccactcttcc tcttcctcca ccactacaac caccgcaaca accaccaaaa
     1861 accctctcaa agaaatttct tttttttctt actttcttgg tttgtcaaat aacagtattc
     1921 agtcgactgg taccaacagg ctccatggct tctaatttta ctcaatttgt tttggttgat
     1981 aatggaggaa ctggagatgt tactgttgct ccttctaatt ttgctaatgg agttgctgaa
     2041 tggatttctt ctaattcaag atctcaggct tataaggtta cttgttctgt tagacaatct
     2101 tctgctcaaa agagaaagta tactattaag gttgaagttc ctaaggttgc tactcaaact
     2161 gttggaggag ttgaattgcc tgttgctgct tggagatctt atttgaatat ggaattgact
     2221 attcctattt ttgctactaa ttctgattgt gaattgattg ttaaggctat gcaaggattg
     2281 ttgaaggatg gaaatcctat tccttctgct attgctgcta attctggaat ttattctgct
     2341 ggaggaggag gatctggagg aggaggatct ggaggaggag gatctggacc taagaagaag
     2401 agaaaggttg ctgctgctgg atcttcaggt ggaagagctg atgctttgga tgattttgat
     2461 ttggatatgt tgggatctga tgctttggat gattttgatt tggatatgtt gggatctgat
     2521 gctttggatg attttgattt ggatatgttg ggatctgatg ctttggatga ttttgatttg
     2581 gatatgttga ttaattaata gctgcaaagc tttcgttcgt atcatcggtt tcgacaacgt
     2641 tcgtcaagtt caatgcatca gtttcattgc gcacacacca gaatcctact gagtttgagt
     2701 attatggcat tgggaaaact gtttttcttg taccatttgt tgtgcttgta atttactgtg
     2761 ttttttattc ggttttcgct atcgaactgt gaaatggaaa tggatggaga agagttaatg
     2821 aatgatatgg tccttttgtt cattctcaaa ttaatattat ttgttttttc tcttatttgt
     2881 tgtgtgttga atttgaaatt ataagagata tgcaaacatt ttgttttgag taaaaatgtg
     2941 tcaaatcgtg gcctctaatg accgaagtta atatgaggag taaaacactt gtagttgtac
     3001 cattatgctt attcactagg caacaaatat attttcagac ctagaaaagc tgcaaatgtt
     3061 actgaataca agtatgtcct cttgtgtttt agacatttat gaactttcct ttatgtaatt
     3121 ttccagaatc cttgtcagat tctaatcatt gctttataat tatagttata ctcatggatt
     3181 tgtagttgag tatgaaaata ttttttaatg cattttatga cttgccaatt gattgacaac
     3241 atgcatcaat cgactatgag taggcaagat gttctgtagg accttgagac cgaaaaggtg
     3301 gtctcagtat tctagaccaa gcatgcacag ctggcgttct cagccagatc cccttgacag
     3361 gatatattgg cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa
     3421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3901 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa
     3961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4201 gtatatatgt gtaacattgg tctagtgatt atttgccgac taccttggtg atctcgcctt
     4261 tcacgtagtg aacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttgtc
     4321 caagataagc ctgcctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     4381 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     4441 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     4501 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     4561 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     4621 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     4681 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     4741 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     4801 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     4861 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     4921 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     4981 gttgagtcga tacttcggcg atcaccgctt ccctcataac accccttgta ttactgttta
     5041 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     5101 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     5161 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     5221 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     5281 tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag cgggcttttt
     5341 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     5401 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     5461 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     5521 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     5581 acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt ttcaattatg
     5641 aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc
     5701 tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc cacaacccgg
     5761 cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa aagcccgaaa
     5821 ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagatctt ggcaggatat
     5881 attgtggtgt aacgttctcg aactaggctc ggacgaagta agcggccgca accttaatta
     5941 aacacggcgc gccaacagcg atcgcaccag gtaccacctc taaaatctgg tttcatatta
     6001 atttcgcaca ccaagttact ttctattatt aactgttata atggaccatg aaatcatttg
     6061 catatgaact gcaatgatac ataatccact ttgttttgtg ggagacattt accagatttc
     6121 ggtaaattgg tattccccct tttatgtgat tggtcattga tcattgttag tggccagaca
     6181 tttgaactcc cgttttttt
//
