LOCUS       sequence-401181-        3873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo level 0 NbGP41-sfGFP-VP64-GB1 coding sequence module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3873)
  AUTHORS   Casas-Mollano JA, Zinselmeier MH, Sychla A, Smanski MJ
  TITLE     Efficient gene activation in plants by the MoonTag programmable 
            transcriptional activator.
  JOURNAL   Nucleic Acids Res. 2023 Jul 21;51(13):7083-7093. doi: 
            10.1093/nar/gkad458.
  PUBMED    37254802
REFERENCE   2  (bases 1 to 3873)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2023-07-21- 21"; volume: "51"; issue: 
            "13"; pages: "7083-7093"
COMMENT     Sequence Label: pMC-J6-NbGP41-sfGFP-VP64-GB1-J9
FEATURES             Location/Qualifiers
     source          1..3873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      111..699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     600..619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     853..870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1344..2054
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1344..2054
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1371..1399)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1994..2014
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2094..2243
                     /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"
     CDS             2268..2288
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2319..2480
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             2508..2558
                     /codon_start=1
                     /product="nuclear localization signal from the human T-cell
                     leukemia virus type 1 (HTLV-1) Rex protein"
                     /label=Rex NLS
                     /translation="PKTRRRPRRSQRKRPPT"
     primer_bind     complement(2610..2628)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2696..2800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3022..3813
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
       61 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      241 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
      421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
      481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
      541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
      661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
      721 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
      781 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
      841 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
      901 cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaatg gaggtgcagc
      961 tggtggaatc tgggggcgga ctggtgcagc ccgggggatc tctgcggctg tcctgcgccg
     1021 cctctggctc catctctagc gtggatgtga tgtcctggta caggcaggcc cctggcaagc
     1081 agagggaact ggtggccttc attactgata ggggaagaac caattacaaa gtgagcgtga
     1141 aaggccgctt cactatcagc cgggataatt ctaagaatat ggtgtatctg cagatgaaca
     1201 gcctgaagcc agaagacact gccgactatc tgtgcagggc tgagtcccgg acttcctggt
     1261 ccagcccatc tcctctggat gtgtgggggc gcggcactca ggtgactgtg tcctccttgg
     1321 atccaggtgg aggtggaagc ggtagcaaag gagaagaact tttcactgga gttgtcccaa
     1381 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg
     1441 aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact ggaaaactac
     1501 ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc ttttcccgtt
     1561 atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac
     1621 aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt
     1681 ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt aaagaagatg
     1741 gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta tacatcacgg
     1801 cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac gttgaagatg
     1861 gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     1921 ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa
     1981 agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca catggcatgg
     2041 atgagctcta caaaggtgga ggtcggaccg gtcgtacggg cggtggcgga ggggatgctt
     2101 tagacgattt tgacttagat atgcttggtt cagacgcgtt agacgacttc gacctagaca
     2161 tgttaggctc agatgcattg gacgacttcg atttagatat gttgggctcc gatgccctag
     2221 atgactttga tctagatatg ctaggtagtg gcggtggcgg aggggctcca aagaagaaga
     2281 gaaaggtccg tacgggagga ggatctcgga ccgaagagta caagcttatc ctgaacggta
     2341 aaaccctgaa aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt
     2401 tcaaacagta cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca
     2461 aaaccttcac ggtaaccgaa ggtggtggta gcggtggtgg tactagtcca aaaacaagga
     2521 ggaggccgcg aagatcacaa cggaaaaggc cgcctacgcc atggccgtaa gctttgagac
     2581 cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2641 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2701 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2761 accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac gcaactggtc
     2821 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     2881 tgactgtttt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     2941 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
     3001 cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta
     3061 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     3121 tacggctccg cagtggatgg cggcctgaag ccacacagcg atattgattt gctggttacg
     3181 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     3241 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     3301 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     3361 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     3421 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     3481 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     3541 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     3601 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     3661 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     3721 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccattacgta
     3781 aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag accaagttta ctcatatata
     3841 ctttagattg atttaaaact tcatttttaa ttt
//
