LOCUS       sequence-321999-        8445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of the MS2 coat protein fused on 
            Ct to the activation domain EDLL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8445)
  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 8445)
  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:EDLL:Tnos (GB1738)
FEATURES             Location/Qualifiers
     source          1..8445
                     /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             1040..1426
                     /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             1433..1453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1457..1477
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1481..1501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1600..1852
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1887..1905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2089..2113
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3414..4043
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4472..5545
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5611..5805
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6041..6063)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6149..6289
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6204..6223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6304..6321)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6475..7063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6555..6574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7151..7966)
                     /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(7151..7966)
                     /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     7874..7893
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8399..8423
                     /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 gatgcccctg 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 taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatgatgg cttctaattt tactcaattt gttcttgttg ataatggtgg
     1081 tactggtgat gttactgttg ctccttctaa ttttgctaat ggtgttgctg aatggatttc
     1141 ttctaattct agatctcaag cttataaggt tacttgttct gttagacaat cttctgctca
     1201 aaatagaaag tatactatta aggttgaagt tcctaaggtt gctactcaaa ctgttggtgg
     1261 tgttgaactt cctgttgctg cttggagatc ttatcttaat atggaactta ctattcctat
     1321 ttttgctact aattctgatt gtgaacttat tgttaaggct atgcaaggtc ttcttaagga
     1381 tggtaatcct attccttctg ctattgctgc taattctggt atttattcgg atcctaaaaa
     1441 gaagcgaaag gtagacccga agaaaaagag gaaggtagat ccaaagaaga agcgcaaggt
     1501 agaagtgttt gaatttgaat atctggatga taaagtgctg gaagaactgc tggatagcga
     1561 agaacgcaaa cgctgagctt ggaatggatc ttcgatcccg atcgttcaaa catttggcaa
     1621 taaagtttct taagattgaa tcctgttgcc ggtcttgcga cgattatcat ataatttctg
     1681 ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg
     1741 gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag
     1801 cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg
     1861 ccaagctaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     1921 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     1981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2041 ctgataaatg cttcaataat gggaccgact cgcgcttgag acgaagcttg acaggatata
     2101 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
     2161 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg
     2221 ggatcaaagt actttgatcc aacccctccg ctgctatagt gcagtcggct tctgacgttc
     2281 agtgcagccg tcatctgaaa acgacatgtc gcacaagtcc taagttacgc gacaggctgc
     2341 cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt agtcgcataa agtagaatac
     2401 ttgcgactag aaccggagac attacgccat gaacaagagc gccgccgctg gcctgctggg
     2461 ctatgcccgc gtcagcaccg acgaccagga cttgaccaac caacgggccg aactgcacgc
     2521 ggccggctgc accaagctgt tttccgagaa gatcaccggc accaggcgcg accgcccgga
     2581 gctggccagg atgcttgacc acctacgccc tggcgacgtt gtgacagtga ccaggctaga
     2641 ccgcctggcc cgcagcaccc gcgacctact ggacattgcc gagcgcatcc aggaggccgg
     2701 cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc accacgccgg ccggccgcat
     2761 ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt tccctaatca tcgaccgcac
     2821 ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag tttggccccc gccctaccct
     2881 caccccggca cagatcgcgc acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa
     2941 agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag
     3001 cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac
     3061 cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg
     3121 caccaggacg gccaggacga accgtttttc attaccgaag agatcgaggc ggagatgatc
     3181 gcggccgggt acgtgttcga gccgcccgcg cacctctcaa ccgtgcggct gcatgaaatc
     3241 ctggccggtt tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa
     3301 accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg
     3361 gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg
     3421 ttatcgctgt acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag
     3481 cccgcgccct gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca
     3541 gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc
     3601 gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg
     3661 gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga
     3721 ttccggtgca gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta
     3781 agcagcgcat tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga
     3841 tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca
     3901 ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa
     3961 ccgttcttga atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg
     4021 aaattaaatc aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa
     4081 cacgctaagt gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct
     4141 ggcagacacg ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa
     4201 gctgaagatg tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat
     4261 cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta
     4321 aaggaggcgg catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt
     4381 gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt tgcctgccgg ccctgcaatg
     4441 gcactggaac ccccaagccc gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt
     4501 acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc
     4561 ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga
     4621 tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc
     4681 gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg
     4741 cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc
     4801 tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc
     4861 cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga
     4921 atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca
     4981 cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct
     5041 ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc
     5101 caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat
     5161 cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta
     5221 ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt
     5281 gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc
     5341 agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa
     5401 gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt
     5461 gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga
     5521 gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc
     5581 aggggaaaaa ggtcgaaaag gactctttcc tgtggatagc acgtacattg ggaacccaaa
     5641 gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg
     5701 gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc
     5761 tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg
     5821 cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc
     5881 cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg
     5941 caatctacca gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca
     6001 aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6061 cggtgacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6121 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     6181 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     6301 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6361 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6421 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6481 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6541 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6601 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6661 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6721 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6781 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6841 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6901 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6961 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7021 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7081 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca
     7141 ttctaggtga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     7201 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     7261 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     7321 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     7381 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     7441 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     7501 ttcgtgattg cgcctgagcg agtcgaaata cgcgatcgct gttaaaagga caattacaaa
     7561 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     7621 aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta
     7681 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     7741 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     7801 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcggtagatt gtcgcacctg
     7861 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     7921 ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcataaca gaacttatta
     7981 tttccttcct cttttctaca gtatttaaag ataccccaag aagctaatta taacaagacg
     8041 aactccaatt cactgttcct tgcattctaa aaccttaaat accagaaaac agctttttca
     8101 aagttgtttt caaagttggc gtataacata gtatcgacgg agccgatttt gaaaccgcgg
     8161 tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc
     8221 atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc ggtaacatga
     8281 gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact gatgggctgc
     8341 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
     8401 gcaggatata ttgtggtgta aacataacaa gcttcgtctc agtca
//
