LOCUS       sequence-310970-        8522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expresion of the viral coat protein MS2 
            fused on C-terminal with the VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8522)
  AUTHORS   Selma S, Bernabe-Orts JM, Vazquez-Vilar M, Diego-Martin B, Ajenjo M,
            Garcia-Carpintero V, Granell A, Orzaez D
  TITLE     Strong gene activation in plants with genome-wide specificity using 
            a new orthogonal CRISPR/Cas9-based programmable transcriptional 
            activator.
  JOURNAL   Plant Biotechnol J. 2019 Sep;17(9):1703-1705. doi: 
            10.1111/pbi.13138. Epub 2019 May 23.
  PUBMED    31034138
REFERENCE   2  (bases 1 to 8522)
  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"; doi: "10.1111/pbi.13138";
            journalName: "Plant Biotechnol J"; date: "2019-09"; volume: "17"; 
            issue: "9"; pages: "1703-1705"
COMMENT     Sequence Label: pDGB3 alpha2 P35S:MS2-VP64:Tnos (GB1395)
FEATURES             Location/Qualifiers
     source          1..8522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1039..1425
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1432..1452
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1456..1476
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1480..1500
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1501..1650
                     /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      1677..1929
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1964..1982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2167..2191
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3492..4121
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4555..5622
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      5688..5882
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6118..6140)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6226..6366
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6281..6300
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6381..6398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6552..7140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6632..6651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7228..8043)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7228..8043)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7951..7970
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8476..8500
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat accacaacaa aacaaacaac aaacaacatt acaatttact
     1021 attctagtcg aaatgatggc ttctaatttt actcaatttg ttcttgttga taatggtggt
     1081 actggtgatg ttactgttgc tccttctaat tttgctaatg gtgttgctga atggatttct
     1141 tctaattcta gatctcaagc ttataaggtt acttgttctg ttagacaatc ttctgctcaa
     1201 aatagaaagt atactattaa ggttgaagtt cctaaggttg ctactcaaac tgttggtggt
     1261 gttgaacttc ctgttgctgc ttggagatct tatcttaata tggaacttac tattcctatt
     1321 tttgctacta attctgattg tgaacttatt gttaaggcta tgcaaggtct tcttaaggat
     1381 ggtaatccta ttccttctgc tattgctgct aattctggta tttattcgga cccaaaaaag
     1441 aagcgtaagg tcgatcccaa gaagaagaga aaggtagatc ctaagaagaa gagaaaggta
     1501 gacgcattgg atgacttcga cttagacatg cttgggtcag atgctttgga cgactttgac
     1561 ctcgacatgt tgggttctga cgcgctagat gacttcgact tggacatgtt gggctccgac
     1621 gctttggacg actttgatct ggacatgtta tgagcttgga atggatcttc gatcccgatc
     1681 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgacga
     1741 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     1801 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     1861 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcdcggtg tcatctatgt
     1921 tactagatcg ggaattgcca agctaattct tgaagacgaa agggcctcgt gatacgccta
     1981 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     2041 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2101 ctcatgagac aataaccctg ataaatgctt caataatggg accgactcgc gcttgagacg
     2161 aagctttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa
     2221 tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca
     2281 accacagggt tcccctcggg atcaaagtac tttgatccaa cccctccgct gctatagtgc
     2341 agtcggcttc tgacgttcag tgcagccgtc atctgaaaac gacatgtcgc acaagtccta
     2401 agttacgcga caggctgccg ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag
     2461 tcgcataaag tagaatactt gcgactagaa ccggagacat tacgccatga acaagagcgc
     2521 cgccgctggc ctgctgggct atgcccgcgt cagcaccgac gaccaggact tgaccaacca
     2581 acgggccgaa ctgcacgcgg ccggctgcac caagctgttt tccgagaaga tcaccggcac
     2641 caggcgcgac cgcccggagc tggccaggat gcttgaccac ctacgccctg gcgacgttgt
     2701 gacagtgacc aggctagacc gcctggcccg cagcacccgc gacctactgg acattgccga
     2761 gcgcatccag gaggccggcg cgggcctgcg tagcctggca gagccgtggg ccgacaccac
     2821 cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc attgccgagt tcgagcgttc
     2881 cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt
     2941 tggcccccgc cctaccctca ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca
     3001 ggaaggccgc accgtgaaag aggcggctgc actgcttggc gtgcatcgct cgaccctgta
     3061 ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag gccaggcggc gcggtgcctt
     3121 ccgtgaggac gcattgaccg aggccgacgc cctggcggcc gccgagaatg aacgccaaga
     3181 ggaacaagca tgaaaccgca ccaggacggc caggacgaac cgtttttcat taccgaagag
     3241 atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca cctctcaacc
     3301 gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc
     3361 agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa
     3421 aacagcttgc gtcatgcggt cgctgcgtat atgatccgat gagtaaataa acaaatacgc
     3481 aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga
     3541 ccatcggaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg
     3601 attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa
     3661 ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg
     3721 acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg
     3781 cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg gccaccgccg
     3841 acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct
     3901 ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg
     3961 ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca
     4021 ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg
     4081 tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa
     4141 aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc
     4201 tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc
     4261 ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga
     4321 gctgctatct gaatagatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta
     4381 gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg
     4441 ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg
     4501 tctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgacggtcgc
     4561 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg
     4621 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg
     4681 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg
     4741 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc
     4801 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg
     4861 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag
     4921 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg
     4981 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc
     5041 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga
     5101 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg
     5161 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg
     5221 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag
     5281 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt
     5341 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc
     5401 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc
     5461 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac
     5521 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc
     5581 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta
     5641 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggac tctttcctgt ggatagcacg
     5701 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag
     5761 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat
     5821 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca
     5881 taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg
     5941 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg
     6001 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac
     6061 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     6121 ctgacacatg cagctcccgg tgacggtcac agcttgtctg taagcggatg ccgggagcag
     6181 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     6241 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     6301 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6361 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6421 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6481 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6541 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6601 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6661 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6721 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6781 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6841 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6901 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6961 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7021 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7081 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7141 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7201 gggattttgg tcatgcattc taggtgatta gaaaaactca tcgagcatca aatgaaactg
     7261 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     7321 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     7381 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     7441 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     7501 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     7561 aaccaaaccg ttattcattc gtgattgcgc ctgagcgagt cgaaatacgc gatcgctgtt
     7621 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     7681 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg
     7741 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     7801 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     7861 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     7921 gtagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     7981 agcatccatg ttggaattta atcgcggcct tgagcaagac gtttcccgtt gaatatggct
     8041 cataacagaa cttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag
     8101 ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac cttaaatacc
     8161 agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta tcgacggagc
     8221 cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac aatcaacatg
     8281 ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa
     8341 tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc tgacgccgtc
     8401 ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag
     8461 ctgttggctg gctggtggca ggatatattg tggtgtaaac ataacaagct tcgtctcagt
     8521 ca
//
